Comparison of the Ca2+-Sensitive Dyes Fluo-3 and Fluo-4 Used with the FLIPR Fluorometric Imaging Plate Reader System
|
|
- Lambert Perry
- 6 years ago
- Views:
Transcription
1 FLIPR Application Note Comparison of the Ca2+-Sensitive Dyes Fluo-3 and Fluo-4 Used with the FLIPR Fluorometric Imaging Plate Reader System INTRODUCTION In the FLIPR System, fluorescence-based measurement of intracellular calcium typically involves the use of the visible spectrum excitable dyes fluo-3, Calcium Green-1 or Oregon Green 488 BAPTA-1. These dyes offer advantages over UVexcitable dyes, including reduced interference from samples and plastic autofluorescence and less cellular photodamage. In addition to those mentioned above, Molecular Probes (Eugene, Oregon) offers an array of visible light-excitable calcium indicators. They differ mainly in their dissociation constant for calcium and in their excitation and emission spectra. However, there is a continued search for dyes with higher sensitivity and lower toxicity. Molecular Probes recently reported the development of a fluo-3 analogfluo-4. The only structural difference between the two dyes are two chlorine atoms in fluo-3 that are replaced by two fluorine atoms in fluo-4. The described beneþts of fluo-4 are greater fluorescence output, faster cell loading, and equivalent fluorescent signal at lower loading concentrations (BioProbes 28, May 1998). This application note describes a direct comparison of results obtained using fluo- 3 and fluo-4, loaded into M 1 -transfected CHO-cells. The FLIPR System was used to measure the calcium response to carbachol activation. MATERIALS AND METHODS and Culture Conditions CHO-M 1 cells, stably transfected with the muscarinic acetylcholine-receptor (M 1 ), were obtained from ATCC (M1 WT3 - CRL1984). The cells were cultured in Ham s F-12 media supplemented with 10% FBS, 2 mm glutamine, Pen/Strep, and 50 µg/ ml Geneticin. were plated one day before the assay on black 96-well microplates with clear bottoms (Costar) at a concentration of 50,000 cells/well (100 µl/well). 1
2 Dye-Loading the Fluo-3 and ßuo-4 (both acetoxymethyl (AM) esters, Molecular Probes) were dissolved in low-water DMSO (puriþed through a molecular sieve) with 10% pluronic acid to a stock concentration of 1 mm. The cells were dye-loaded in regular culture medium (see above) that also included ßuo-3 or ßuo-4 at the concentrations given in Table 1. To prevent dye extrusion by the anion exchange protein, the loading medium was supplemented with 2.5 mm probenicid, and 20 mm HEPES, ph 7.3, was added to maintain optimal ph. To compare the loading efþcacy of ßuo-3 and ßuo-4, dye concentration and loading time were varied from those in a standard loading procedure* (2 µm ßuo-3, 1.0 hour loading). After loading, a Labsystems Cellwash 4 was used to wash the cells three times with HBSS containing 20 mm HEPES and 2.5 mm Probenicid. After the last wash step, 100 µl buffer per well was left in the microplate. The conditions deþned in this investigation are not universally optimal. Optimal conditions will depend on the speciþc application and will be affected by many factors, e.g. cell type, receptor type, expression status of the receptor, and coupling of the receptor. Therefore, some users might need to apply higher dye concentrations to gain higher loading intensities and signals in their assays. Fluorescent Dye Concentration Loading time fluo-3 2 µm 0.5 hour fluo-3* 2 µm* 1.0 hour* fluo-4 1 µm 0.5 hour fluo-4 1 µm 1.0 hour fluo-4 2 µm 0.5 hour fluo-4 2 µm 1.0 hour Table 1: Evaluated loading conditions (* = standard condition). Agonist Stimulation of In the FLIPR system, 50 µl aliqouts of increasing concentrations of the agonist carbachol (at 3x their Þnal concentration) were added to the cell plate at 50 µl/sec. The Þnal concentrations in the wells were: 2.5 µm, 1.25 µm, 625 nm, 312 nm, 156 nm, 78 nm, 39 nm, 19 nm, and 10 nm. The FLIPR system was set for 0.4 sec exposure time, F-stop of F/2 and laser power of 400 mw, and the change in ßuorescence was monitored before, during, and after the carbachol addition. RESULTS of Non-Loaded (Table 2, Column 1) The average background of non-loaded cells was about 1,450 counts (n = 8). This is due to the autoßuorescence of the plastic and the intrinsic ßuorescence of the cells, which mainly comes from the ßavin coenzymes FAD and FMN: Flavin-Adenine-Dinucleotide and Flavin-Mononucleotide, whose absorption/emission wavelengths are about 450/515 nm (J. Histochem Cytochem 27:44 [1979]; J. Histochem Cytochem 27:36 [1979]). 2
3 of Loaded (Table 2, Columns 2 and 3) The background of loaded cells varied with different loading conditions. Lengthening the loading time for either ßuo-3 or ßuo-4 from 0.5 hour to 1.0 hour increased the basal ßuorescence intensity (indicated by an increase in basal counts). Increasing the dye concentration from 1 µm to 2 µm also increased the basal ßuorescence intensity of ßuo-4 (not tried with ßuo-3). For the conditions tested, not surprisingly, the basal ßuorescence intensity increased with increasing dye concentration and increasing loading time. Maximal Response After Stimulation (Table 2, Columns 4 and 5) The two highest concentrations of carbachol (1.25 µm and 2.5 µm) both led to a maximal stimulation of the cells as measured by calcium efßux. Therefore the average ßuorescent counts from the cells stimulated with these two saturating concentrations of carbachol was taken as the maximal response. exposed to 1 µm ßuo-4 for a 0.5 hour loading time gave a robust signal of 10,994 counts above basal ßuorescence when stimulated. Doubling the incubation time to 1.0 hour and the concentration to 2 µm signiþcantly increased the signal to 24,289 counts. Increasing the loading time for ßuo-3 also resulted in higher signal when stimulated (0.5 hour: 6525 counts; 1.0 hour: 9765 counts). In summary, increasing the dye concentration or loading time resulted in an increase in both the background and the stimulated signal. Induction Level of the Signal (Table 2, Column 6) To estimate the induction level of calcium upon stimulation, the corrected response of maximally stimulated cells (column 5) was divided by the corrected background of non-stimulated, loaded cells (column 3). By comparing 0.5 hour loading time with 1.0 hour loading time (with a Þxed dye concentration) one can see that ßuo-3 (2 µm) and ßuo-4 (1 µm and 2 µm) produce a higher degree of induction with a shorter incubation time ( / / ). By comparing the response to 1 µm and 2 µm ßuo-4 (with a Þxed loading time) it also appears that a higher dye concentration also leads to a higher degree of induction. By comparing ßuo-3 with ßuo-4 under the same loading conditions (both 2 µm and 0.5 hour or 1.0 hour loading), it appears that the ßuo-4 dye produces a higher magnitude of induction. 3
4 Dye Concent. Loading Time fluo-3 2 µm 0.5 hr fluo-3* 2 µm* 1.0 hr* fluo-4 1 µm 0.5 hr fluo-4 1 µm 1.0 hr fluo-4 2 µm 0.5 hr fluo-4 2 µm 1.0 hr Unloaded 1,469 +/- 72 1,469 +/- 72 1,415 +/- 58 1,458 +/- 69 1,415 +/- 58 1,458 +/- 69 (2) Loaded (3) Loaded correct. for 3,580 +/ ,111 5,158 +/ ,689 4,857 +/ ,442 7,355 +/ ,897 5,533 +/ ,118 9,909 +/ ,451 (4) Stimulated (5) Stimulated correct. for 7,994 +/-532 6,525 11,234 +/ ,765 12,409 +/ ,994 17,721 +/- 1,360 16,263 16,242 +/ ,827 25,747 +/- 1,814 24,289 (6) Induction Level = (5)/(3) Table 2: CHO-M 1 loaded with ßuo-3 (variable loading time) or ßuo-4 (variable loading time and dye concentration). In columns to (5) ßuorescent counts are given. * = Standard conditions from unloaded cells and plastic; n = 8 (2) Dye loaded, unstimulated cells; n = 44 (3) Dye loaded, unstimulated cells, corrected for background ßuorescence (4) Maximally (1.25 or 2.5 µm carbachol) stimulated cells; n = 8 (5) Maximally (1.25 or 2.5 µm carbachol) stimulated cells, corrected for background (6) Induction level= (5)/(3) COMPARISON OF DOSE RESPONSE CURVES Figure 1 shows the dose response curves generated for the 6 different loading conditions (values expressed as a percentage of the maximum for each individual condition). As expected, the response curves are close together and neither different dyes nor different loading conditions signiþcantly change them (and therefore the corresponding EC 50 values are also unchanged, remaining between 70 and 95 nm). Fluorescent Change (% of Max.) µm fluo-3; 0.5 hr 2 µm fluo-3; 1.0 hr 1 µm fluo-4; 0.5 hr 1 µm fluo-4; 1.0 hr 2 µm fluo-4; 0.5 hr 2 µm fluo-4; 1.0 hr Carbachol (nm) Figure 1: Dose response curves for different dyes, dye concentrations, and loading times (n = 4). 4
5 DISCUSSION These results show that fluo-4 may offer several advantages over the standardly used fluo-3. When using 2 µm fluo-4, the loading time can be decreased from 1.0 hour to 0.5 hour with equivalent or better loading than that seen with fluo-3. Decreased loading times are beneþcial not only because they save time, but also because the reduced exposure time may also decrease potential toxic effects of the calcium-sensitive dye. In addition, shorter loading times reduce the amount of dye that moves from the cytoplasm into intracellular compartments (e.g. mitochondria). The decreased loading times possible when using fluo-4 also improves the signaltonoise ratio by increasing the ratio of the fluorescence produced by stimulated versus unstimulated cells. This could be because decreased dye exposure may be more favorable to cell viability. The reduced relative background fluorescence may also be attributable to a reduction in the amount of dye accumulated in the intracellular compartments. This study has been performed using a CHO cell line, which is frequently used in HTS and in the FLIPR system. However, dyes behave differently in different cell lines and it is important that appropriate dye and loading conditions are tested for each new cell line being studied. SALES OFFICES United States & Canada Molecular Devices Tel Fax China Molecular Devices Beijing Tel Fax Germany Molecular Devices GmbH Tel / Fax / South Korea Molecular Devices Korea, LLC Tel Fax FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. The trademarks used herein are the property of Molecular Devices, Inc. or their respective owners. Specifications subject to change without notice Molecular Devices, Inc. Printed in U.S.A. 6/10 # B Brazil Molecular Devices Brazil Tel Fax Molecular Devices Shanghai Tel Fax Japan Molecular Devices Japan, Osaka Tel Fax United Kingdom Molecular Devices Ltd. Tel Fax Molecular Devices Japan, Tokyo Tel Fax
FLIPR Calcium 6 Assay Explorer Kit (R8190) FLIPR Calcium 6-QF Assay Explorer Kit (R8192)
FLIPR Calcium 6 Assay Explorer Kit (R8190) FLIPR Calcium 6-QF Assay Explorer Kit (R8192) About the FLIPR Calcium 6 and 6-QF Assay Kits The FLIPR Calcium 6 Assay Kits from Molecular Devices, LLC (hereafter
More informationFluo-4 NW Calcium Assay Kits (F36205, F36206)
Product Information Revised: 20 October 2006 Fluo-4 NW Calcium Assay Kits (F36205, F36206) F36205 Fluo-4 NW Calcium Assay Kit (high-throughput) *for 100 microplates* F36206 Fluo-4 NW Calcium Assay Kit
More informationRhod-4 No Wash Calcium Assay Kit
ab112157 Rhod-4 No Wash Calcium Assay Kit Instructions for Use For detecting calcium in cells without washing by using our proprietary fluorescence probe This product is for research use only and is not
More informationFluo-8 Medium Removal Calcium Assay Kit
ab112128 Fluo-8 Medium Removal Calcium Assay Kit Instructions for Use For detecting calcium in cells by using our proprietary fluorescence probe This product is for research use only and is not intended
More informationMeasuring Membrane Potential using the FLIPR Membrane Potential Assay Kit on Fluorometric Imaging Plate Reader (FLIPR ) Systems
FLIPR Application Note Measuring Membrane Potential using the FLIPR Membrane Potential Assay Kit on Fluorometric Imaging Plate Reader (FLIPR ) Systems 2 Table of Contents Section I: Introduction..........................................3
More informationFluo-8 No Wash Calcium Assay Kit
ab112129 Fluo-8 No Wash Calcium Assay Kit Instructions for Use For detecting calcium in cells by using our proprietary fluorescence probe This product is for research use only and is not intended for diagnostic
More informationab Fluo-8 No Wash Calcium Assay Kit
ab112129 Fluo-8 No Wash Calcium Assay Kit Instructions for Use For detecting calcium in cells by using our proprietary fluorescence probe. This product is for research use only and is not intended for
More informationGPCR induction followed by a fluorometric Ca 2+ assay in Thermo Scientific Varioskan LUX
GPCR induction followed by a fluorometric Ca 2+ assay in Thermo Scientific Varioskan LUX SP&A Application Laboratory, Thermo Fisher Scientific, Vantaa, Finland Application Notes Key Words Cellular signal,
More informationFLIPR Calcium 3 Assay Kit Product # R8091 (Explorer Kit) R8090 (Bulk Kit) R8108 (Express Kit)
Product Insert FLIPR Calcium 3 Assay Kit Product # R8091 (Explorer Kit) R8090 (Bulk Kit) R8108 (Express Kit) Introduction About the FLIPR Calcium 3 Assay Kit The Calcium 3 Assay Kit from Molecular Devices
More informationCalcium Assay Kit. Technical Data Sheet. Product Information. Description. Storage. Materials not included
BD Technical Data Sheet Calcium Assay Kit Product Information Catalog Number: 640176 Size Reagents for 10 plates Components: Calcium Indicator, 1 vial, lyophilized 10X Signal Enhancer, 10 ml 1X Calcium
More informationIntroduction. Application Protocol. FlexStation Systems
FlexStation Systems Optimization of a muscarinic M 3 -receptor assay using frozen Assay Ready CHO cells on the FlexStation 3 System: The impact of calcium dye selection and cell culture conditions By Simon
More informationFaster Chemiluminescent camp detection by HitHunter camp XS+ on the SpectraMax L Luminescence Microplate Reader
SpectraMax Systems Faster Chemiluminescent camp detection by HitHunter camp XS+ on the SpectraMax L Luminescence Microplate Reader By Rajini Bompelli and Sunitha Ambikapathi, DiscoveRx Corporation, 42501
More informationRatiometric Calcium Assay Kit
BD Technical Data Sheet Ratiometric Calcium Assay Kit Product Information Catalog Number: 644244 Components: Ratiometric Calcium Indicator, 10 vials, lyophilized 10X Signal Enhancer, 100 ml Description
More informationFLIPR Potassium Assay Kit
FLIPR Potassium Assay Kit Table A-1: Available Kits Assay Kit Explorer Kit Bulk Kit FLIPR Potassium Assay Kit R8222 R8223 The FLIPR Potassium Assay Kit from Molecular Devices provides a fast, simple, and
More informationab Fura-2 No Wash Calcium Assay Kit
ab176766 Fura-2 No Wash Calcium Assay Kit Instructions for use: For the measurement of intracellular calcium mobilization in live cells. This product is for research use only and is not intended for diagnostic
More informationBD Ratiometric Calcium Assay Kit
BD Technical Data Sheet BD Ratiometric Calcium Assay Kit Product Information Catalog Number: 644243 Components: Ratiometric Calcium Indicator, 1 vial, lyophilized 10X Signal Enhancer, 10 ml Calcium Assay
More informationCell Viability Assay Kit Fluorometric Near InfraRed
ab112123 Cell Viability Assay Kit Fluorometric Near InfraRed Instructions for Use For detecting cell viability in suspension and adherent cells by using Near InfraRed fluorescence probes This product is
More informationA Cost-effective Workflow for High-Throughput Screening of G- Protein Coupled Receptors (GPCRs)
A p p l i c a t i o n N o t e A Cost-effective Workflow for High-Throughput Screening of G- Protein Coupled Receptors (GPCRs) Monitoring Receptor Mediated Calcium Flux Paul Held and Peter Banks, BioTek
More informationab MDR Assay Kit (Fluorometric)
ab112142 MDR Assay Kit (Fluorometric) Instructions for Use For detecting MDR pump activities in cells using our proprietary fluorescence probe. This product is for research use only and is not intended
More informationEarlyTox Cardiotoxicity Kit
EarlyTox Cardiotoxicity Kit Table 1-1: Available Kits Assay Kit Explorer Kit Bulk Kit EarlyTox Cardiotoxicity Kit R8210 R8211 The EarlyTox Cardiotoxicity Kit from Molecular Devices provides a fast, simple,
More informationab Cell Viability Assay Kit Fluorometric Dual Green/Red
ab112121 Cell Viability Assay Kit Fluorometric Dual Green/Red Instructions for Use For detecting cell viability in suspension and adherent cells by using dual proprietary green and red fluorescence probes.
More informationCell Viability Assay Kit Fluorometric Blue
ab112120 Cell Viability Assay Kit Fluorometric Blue Instructions for Use For detecting cell viability in suspension and adherent cells by using our proprietary Blue fluorescence probe This product is for
More informationFluo Calcium Indicators
Product Information Revised: 26 November 2001 Storage upon receipt: Indicator salts Room temperature, 4 C or 20 C $ Desiccate $ Protect from light Dextran conjugate 20 C $ Desiccate $ Protect from light
More informationFlexStation 3 Microplate Reader
FlexStation 3 Microplate Reader A five-mode microplate reader with integrated fluid transfer > five-mode reader with wide range of applications > flexible liquid transfer enables more assay conditions
More informationMembrane Potential Assays Using the ValiScreen Human Kv1.3 Voltage-Gated K + Channel Cell Line on the EnVision Multilabel Plate Reader
TECHNICAL BRIEF ValiScreen Stable Recombinant Ion Channel Cell Line and the EnVision Multilabel Plate Reader Membrane Potential Assays Using the ValiScreen Human Kv1.3 Voltage-Gated K + Channel Cell Line
More informationRhod-3 Calcium Imaging Kit
Rhod-3 Calcium Imaging Kit Catalog no. R10145 Table 1. Contents and storage information. Material Amount Storage Stability Rhod-3, AM (Component A); MW ~1600 1 vial 2 C 8 C Protect from light Dimethylsulfoxide
More informationhuman Muscarinic Acetylcholine Receptor M 4 Aequorin Cell Line
TECHNICAL DATA SHEET AequoScreen GPCR Cell Line Caution: For Laboratory Use. A research reagent for research purposes only human Muscarinic Acetylcholine Receptor M 4 Aequorin Cell Line Product No.: ES-213-A
More informationab JC1 - Mitochondrial Membrane Potential Assay Kit
ab113850 JC1 - Mitochondrial Membrane Potential Assay Kit Instructions for Use For the measurement of mitochondrial membrane potential by fluorescence plate reader This product is for research use only
More informationFlexStation 3 microplate reader
FlexStation 3 microplate reader A five-mode microplate reader with integrated fluid transfer > five-mode reader with wide range of applications > flexible liquid transfer enables more assay conditions
More informationPyrophosphate Assay Kit (Fluorometric)
ab112155 Pyrophosphate Assay Kit (Fluorometric) Instructions for Use For the measurement of Pyrophosphate using our proprietary fluorescence probe for screening inhibition or enzyme activity This product
More informationEarlyTox Cell Integrity Kit
EarlyTox Cell Integrity Kit The EarlyTox Cell Integrity Kit from Molecular Devices is an optimized set of reagents that simplifies the measurement of live and dead cells in a single well. The assay uses
More informationApplication Note INTRODUCTION
Application Note 20 A microtiter-based assay for the determination of ID 50 s of β-lactamase inhibitors employing reporter substrates detected at UV or visible wavelengths INTRODUCTION β-lactamases are
More informationEnzChek Phospholipase A 2 Assay Kit
EnzChek Phospholipase A 2 Assay Kit Catalog nos. E10217, E10218 Table 1. Contents and storage information. Material E10217 (2-plates) E10218 (10-plates) Concentration Storage Stability Phospholipase A
More informationab CytoPainter Mitochondrial Staining Kit NIR Fluorescence
ab176747 CytoPainter Mitochondrial Staining Kit NIR Fluorescence Instructions for Use For staining Mitochondria in live cells with our proprietary NIR probe. This product is for research use only and is
More informationhuman Muscarinic Acetylcholine Receptor M 3 Cell Line
TECHNICAL DATA SHEET ValiScreen GPCR Cell Line Caution: For Laboratory Use. A research product for research purposes only human Muscarinic Acetylcholine Receptor M 3 Cell Line Product No.: ES-212-C Lot
More informationPerformance of cell viability and cytotoxicity assays on the IN Cell Analyzer 3000
GE Healthcare Application Note 28-4070-51 AA IN Cell Analyzer 3000 Performance of cell viability and cytotoxicity assays on the IN Cell Analyzer 3000 Key words: cell-based assay viability cytotoxicity
More informationab CytoPainter Live Cell Labeling Kit - Blue Fluorescence Instructions for Use
ab187963 CytoPainter Live Cell Labeling Kit - Blue Fluorescence Instructions for Use For labelling live cells in blue fluorescence for the studies that require the fluorescent tag molecules retained inside
More informationContaminant human transferrin assay
ILA Application Note Contaminant human transferrin assay INTRODUCTION Human transferrin is an 8, Dalton glycoprotein found in human serum that facilitates transport of iron between cells. The iron-poor
More informationab CytoPainter Fixable Cell Viability Assay Kit (Fluorometric - Green Ex 405 nm)
ab176739 - CytoPainter Fixable Cell Viability Assay Kit (Fluorometric - Green Ex 405 nm) Instructions for Use For evaluation of the viability of mammalian cells by fluorescent labeling and flow cytometry.
More informationab CytoPainter Fixable Cell Viability Assay Kit (Fluorometric - Blue Ex 405 nm)
ab176738 - CytoPainter Fixable Cell Viability Assay Kit (Fluorometric - Blue Ex 405 nm) Instructions for Use For evaluation of the viability of mammalian cells by fluorescent labeling and flow cytometry.
More informationMeasuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers
APPLICATION NOTE Measuring cell proliferation using the CyQUANT Cell Proliferation Assay with SpectraMax Microplate Readers Introduction Quantitation of cell proliferation using fluorescence allows one
More informationApplications of rapid immunoassays (FAST ILA) in cell culture/fermentation and process development using the Threshold System
ILA Application Note Applications of rapid immunoassays (FAST ILA) in cell culture/fermentation and process development using the Threshold System INTRODUCTION Accurate measurement of the concentration
More informationhuman Vasopressin V 1A Receptor Cell Line
TECHNICAL DATA SHEET ValiScreen GPCR Cell Line Caution: For Laboratory Use. A research product for research purposes only human Vasopressin V 1A Receptor Cell Line Product No.: ES-361-C Lot No.: M1W-C1
More informationDNAse I qualification and sample treatment
DNA Application Note DNAse I qualification and sample treatment INTRODUCTION The Threshold Total DNA Assay quantitates contaminant DNA in biopharmaceutical drugs. DNase I can be used in conjunction with
More informationFor the rapid, sensitive and accurate measurement of Lactate levels in various samples
ab65330 L Lactate Assay Kit (Fluorometric) Instructions for Use For the rapid, sensitive and accurate measurement of Lactate levels in various samples This product is for research use only and is not intended
More informationTime-Resolved Fluorescence Based GTP Binding Assay for G-Protein Coupled Receptors
Time-Resolved Fluorescence Based GTP Binding Assay for G-Protein Coupled Receptors Gregory Warner, Rita Syystö, Heini Frang, Christel Gripenberg-Lerche, Jurgen Vanhauwe, Tarja Ahola, and Satu Kovanen Abstract
More informationCytoPainter Cell Tracking Staining Kit Green Fluorescence
ab138891 CytoPainter Cell Tracking Staining Kit Green Fluorescence Instructions for Use For labelling live cells in green fluorescence for the studies that require the fluorescent tag molecules retained
More informationCytoPainter Cell Tracking Staining Kit Green Fluorescence
ab138891 CytoPainter Cell Tracking Staining Kit Green Fluorescence Instructions for Use For labelling live cells in green fluorescence for the studies that require the fluorescent tag molecules retained
More informationTotal NAD and NADH Assay Kit (Colorimetric)
ab186032 Total NAD and NADH Assay Kit (Colorimetric) Instructions for Use An optimized assay for monitoring Total NAD and NADH. This product is for research use only and is not intended for diagnostic
More informationAutomated Imaging Assay for Characterizing Ca 2+ Flux with R-GECO Biosensor
A p p l i c a t i o n N o t e Automated Imaging Assay for Characterizing Ca 2+ Flux with R-GECO Biosensor Joe Clayton, Ph.D., and Peter Banks, Ph.D., BioTek Instruments, Inc., Winooski, VT USA Abstract
More information- coupled receptors in either adherent or suspension cells.
IP-One - Gq KIT PROTOCOL IP-ONE - Gq KIT: Part #: 62IPAPEB (1,000 tests), 62IPAPEC (20,000 tests), 62IPAPEJ (100,000 tests) Version: #6 (Feb 2017) Storage temperature: 2-8 C For research use only. Not
More informationCalcein AM Cell Viability Kit
Instructions For Research Use Only. Not For Use In Diagnostic Procedures Calcein AM Cell Viability Kit Catalog# 4892-010-K 1000 Tests* * Calculated based on using 1 μm final concentration of Calcein AM;
More informationApplication Note. Developed for: Aerius, Odyssey Classic, Odyssey CLx, and Odyssey Sa Infrared Imaging Systems
Application Note On-Cell Western Plate-Based Assay for Targeted Near-Infrared-Labeled Optical Imaging Agent Development: Receptor-Based Binding and Competition Assays Developed for: Aerius, Odyssey Classic,
More informationIntracellular ph (phi) Detection
Intracellular ph (phi) Detection Table 1 Intracellular ph (phi) detection products phrodo Red AM Intracellular ph Indicator (Cat. no. P35372) Amount Concentration Storage phrodo Red AM Intracellular ph
More informationab Proteasome 20S Activity Assay Kit (Fluorometric)
ab112154 Proteasome 20S Activity Assay Kit (Fluorometric) Instructions for Use For detecting proteasome activity in cultured cells using our proprietary green fluorescence probe. This product is for research
More informationA Comparison of PerkinElmer s LANCE camp Assay Performance to that of State of the Art Competitive Technologies
A Comparison of PerkinElmer s LANCE camp Assay Performance to that of State of the Art Competitive Technologies GPCR ASSAYS A P P L I C A T I O N N O T E Introduction G-protein coupled receptors (GPCRs)
More informationhuman Serotonin 5-HT 2A Receptor Cell Line
TECHNICAL DATA SHEET ValiScreen GPCR Cell Line Caution: For Laboratory Use. A research product for research purposes only human Serotonin 5-HT 2A Receptor Cell Line Product No.: ES-313-C Lot No.: M1W-C3
More informationab JC-10 Mitochondrial Membrane Potential Assay Kit Flow Cytometry
ab112133 JC-10 Mitochondrial Membrane Potential Assay Kit Flow Instructions for Use For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe. This product
More informationFast and Easy Calcium Flux Assays on the VICTOR Nivo Multimode Plate Reader
APPLICATION NOTE Multimode Detection Authors: Adriana-Maria Stanciu Maria Kuzikov Fraunhofer IME Screening Port Hamburg, Germany Fast and Easy Calcium Flux Assays on the VICTOR Nivo Multimode Plate Reader
More informationMeasuring KCNT1 Ion Channel Activities Using the FluxOR Potassium Ion Channel Assay Kit on the VICTOR Nivo Multimode Reader
APPLICATION NOTE Multimode Detection Authors: Ebru Betül Yurtseven Maria Kuzikov Fraunhofer IME ScreeningPort, Germany Measuring KCNT1 Ion Channel Activities Using the FluxOR Potassium Ion Channel Assay
More informationTag-lite Tachykinin NK1 labeled Cells, ready-to-use (transformed & labeled), 200 tests* (Part# C1TT1NK1)
TAG - L I T E Tag-lite Tachykinin NK1 Receptor Ligand Binding Assay protocol Tachykinin NK1 labeled cells for: 200 tests Part#: C1TT1NK1 Lot#: 03-1 March 2016 (See expiration date on package label) Store
More informationAdvanced phospho-erk1/2 (Thr202/Tyr204) 500 tests
Headquarters & Europe Office Cisbio Bioassays Phone: +33 (0)4 66 79 67 05 Fax: +33 (0)4 66 79 19 20 bioassays@cisbio.com cisbio_dd_pi_64aerpeg USA Office Cisbio US, Inc. Phone: +1 888 963 4567 Fax: +1
More informationNew and Highly Sensitive AlphaScreen camp Assay to Measure Femtomol camp Variations in Cell and Membrane Based Assays
New and Highly Sensitive AlphaScreen camp Assay to Measure Femtomol camp Variations in Cell and Membrane Based Assays 1 Illy, C., 1 Boissonneault, M., 1 Bouchard, N., 2 Dimeo, J., 1 Hamann, G., 1 Legault,
More informationhuman Angiotensin AT 2 Receptor Cell Line
TECHNICAL DATA SHEET ValiScreen GPCR Cell Line Caution: For Laboratory Use. A research product for research purposes only human Angiotensin AT 2 Receptor Cell Line Product No.: ES-070-C Lot No.: M1W-C1
More informationAccelerating Scientific Discovery
Elite TM Fluorescent Membrane Potential Dye Kit CATALOG NUMBER: CA-M165 Description camp is a key second messenger involved extensively in cellular signal transduction pathways associated with the majority
More informationSensoLyte Homogeneous AFC Caspase-3/7 Assay Kit
SensoLyte Homogeneous AFC Caspase-3/7 Assay Kit Catalog # 71114 Unit Size Kit Size 1 Kit 500 Assays (96-well) or 1250 Assays (384-well) This kit is optimized to detect caspase-3/7 activity in cell culture
More informationUniversal Kinase Assay Kit (Fluorometric) For monitoring ADP formation, which is directly proportional to enzyme phosphotransferase activity
ab138879 Universal Kinase Assay Kit (Fluorometric) Instructions for Use For monitoring ADP formation, which is directly proportional to enzyme phosphotransferase activity This product is for research use
More informationThiolTracker Violet (Glutathione Detection Reagent)
ThiolTracker Violet (Glutathione Detection Reagent) Catalog nos. T10095, T10096 Table 1. Contents and storage information. Material Amount Concentration Storage* Stability ThiolTracker Violet 180 assays
More informationHuman Opioid mu (OP3) Receptor, Frozen Cells
TECHNICAL DATA SHEET campzen Caution: For Laboratory Use. A research reagent for research purposes only. You are authorized to utilize these frozen cell preparations one time only. Any attempt to transfer,
More informationXanthine Oxidase Assay Kit
ab102522 Xanthine Oxidase Assay Kit Instructions for Use For the rapid, sensitive and accurate measurement of Xanthine Oxidase activity in various samples. This product is for research use only and is
More informationViolet Chromatin Condensation/Dead Cell Apoptosis Kit with Vybrant DyeCycle Violet and SYTOX AADvanced for Flow Cytometry
Violet Chromatin Condensation/Dead Cell poptosis Kit with Vybrant DyeCycle Violet and SYTOX Dvanced for Flow Cytometry Catalog no. 35135 Table 1. Contents and storage information. Material mount Concentration
More informationApplication Note LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SPECTRAmax GEMINI XS Fluorescence Microplate Reader
Application Note 43 LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SPECTRAmax GEMINI XS Fluorescence Microplate Reader Anne T. Ferguson, Ph.D. Molecular Devices Corporation 1 2 INTRODUCTION
More informationTranscreener PDE Assay
Technical Manual Transcreener PDE Assay Transcreener PDE Assay Kit Catalog #3006-1K, 3006-10K 1.0 Introduction p.2 2.0 Assay Principle p.2 3.0 Instrumentation p.3 4.0 Transcreener PDE Assay Kit Components
More informationDownloaded from and current as of 02/01/2008
Downloaded from http://www.moleculardevices.com and current as of // Veritas microdissection system: optimized protocol for laser microdissection of living in vitro cells Veritas application Note By Steven
More informationAccelerate cell line development. Get to your high value hits faster
Accelerate cell line development Get to your high value hits faster Transform your workflows to elevate productivity Fast screening and selection of secretory cell lines with ClonePix Systems Screen 10x
More informationSensoLyte Homogeneous AFC Caspase-8 Assay Kit *Fluorimetric*
SensoLyte Homogeneous AFC Caspase-8 Assay Kit *Fluorimetric* Catalog # 7288-2 Kit Size 2 Assays (96-well plate) Optimized Performance: This kit is optimized to detect caspase-8 activity. Enhanced Value:
More informationab Autophagy/ Cytotoxicity Dual Staining Kit
ab133075 Autophagy/ Cytotoxicity Dual Staining Kit Instructions for Use For the study of the regulation of autophagy and cytotoxicity at the cellular level. This product is for research use only and is
More informationMeasure-iT Thiol Assay Kit (M30550)
Product Information Revised: 24 March 2008 Measure-iT Thiol Assay Kit (M30550) Quick Facts Storage upon receipt: Store at 20ºC Protect from light Desiccate Avoid freeze/thaw cycles thiol quantitation buffer
More informationAsante NaTRIUM Green 1 (ANG-1)
Page 1 of 10 Asante NaTRIUM Green 1 (ANG-1) ANG-1 is to Sodium as Fluo-4 is to Calcium ANG-1 Highlights Compatible with green argon laser, LED excitation, or two-photon IR laser excitation Twenty-fold
More informationApplication Note 2011/06. Orion II Microplate Luminometer Simplicity 4
Berthold Detection Systems GmbH Bleichstrasse 56 68 D-75173 Pforzheim/Germany Phone: +49(0)7231/9206-0 Fax: +49(0)7231/9206-50 E-Mail: contact@titertek-berthold.com Internet: www.titertek-berthold.com
More informationHuman Muscarinic M2 Receptor, -Irradiated Frozen Cells
TECHNICAL DATA SHEET AequoZen Caution: For Laboratory Use. A research reagent for research purposes only Human Muscarinic M2 Receptor, -Irradiated Frozen Cells Product No.: ES-211-AF Lot No.: 639-110-A
More informationab Generic Caspase Activity Assay Kit Fluorometric Green
ab112130 Generic Caspase Activity Assay Kit Fluorometric Green Instructions for Use For detecting Caspase activity in cells by using our proprietary green fluorescence probe. This product is for research
More informationService solutions for peak performance. Flexible service plans, maximum productivity, expert engineers.
Service solutions for peak performance Flexible service plans, maximum productivity, expert engineers www.moleculardevices.com Choose the right service plan for your specific laboratory priorities Performance
More informationLow Background D-A-D Type Fluorescent Probe for Imaging of Biothiols
Electronic Supplementary Material (ESI) for Journal of Materials Chemistry B. This journal is The Royal Society of Chemistry 2018 Electronic supplementary information Low Background D-A-D Type Fluorescent
More informationCELL HEALTH. reliable cell viability data. instant time savings. PrestoBlue Cell Viability Reagent
CELL HEALTH reliable cell viability data instant time savings PrestoBlue Cell Viability Reagent PrestoBlue Cell Viability Reagent Why should I use the PrestoBlue Cell Viability Reagent? PrestoBlue reagent
More informationContaminant bovine transferrin assay
ILA Application Note Contaminant bovine transferrin assay INTRODUCTION Bovine transferrin, a 76, Dalton glycoprotein, is one of the constituents of bovine serum. Transferrin found in serum can be associated
More informationAmplex UltraRed Reagent
Amplex UltraRed Reagent Catalog no. A366 Table 1. Contents and storage information. Material Amount Storage Stability Amplex UltraRed reagent (MW = ~3) 5 vials, each containing 1 mg 2 C Desiccate Protect
More informationab CytoPainter Mitochondrial Staining Kit - Green Fluorescence
ab112143 CytoPainter Mitochondrial Staining Kit - Green Fluorescence Instructions for Use For staining Mitochondria in live cells using our proprietary green fluorescence probe This product is for research
More informationFor the rapid, sensitive and accurate measurement of Creatinine in various samples.
ab65340 Creatinine Assay Kit Instructions for Use For the rapid, sensitive and accurate measurement of Creatinine in various samples. This product is for research use only and is not intended for diagnostic
More informationApoptosis And Anti-tumor Effect Induced By Mtor Inhibitor And Autophagy Inhibitor In Human Osteosarcoma Cells
Apoptosis And Anti-tumor Effect Induced By Mtor Inhibitor And Autophagy Inhibitor In Human Osteosarcoma Cells Ryosuke Horie. Kagawa University of medecine, Kita-gun, Japan. Disclosures: R. Horie: None.
More informationSensoLyte 570 Generic MMP Assay Kit *Fluorimetric*
SensoLyte 570 Generic MMP Assay Kit *Fluorimetric* Catalog # 72101 Kit Size 100 Assays (96-well plate) Optimized Performance: Optimal conditions for detection of generic MMP activity Enhanced Value: Less
More informationFunctional GPCR Studies Using AlphaScreen camp Detection Kit
PPLICTION NOTE lpha Technology uthors: Jen Carlstrom Dawn Nida PerkinElmer, Inc. Hopkinton, M Functional GPCR Studies Using lphascreen cmp Detection Kit Introduction G protein-coupled receptors (GPCRs)
More informationCytotoxicity LDH Assay Kit-WST
Cytotoxicity LDH Assay Kit-WST Supplementary Information Notice to Users This instruction complements the Technical Manual in the product. Please use this instruction as supplements of the Technical Manual.
More informationMeasurement of Green Fluorescent Protein in the SPECTRAmax GEMINI-XS Spectrofluorometer.
Application Note 44 Measurement of Green Fluorescent Protein in the SPECTRAmax GEMINI-XS Spectrofluorometer. Simon Lydford 1 and Thomas Giller 2 1 Molecular Devices Limited and 2 Axovan Ltd. INTRODUCTION
More information+ M III. IMAP Screening Express Kit Product #8073 Quantity: 8000, 20 µl reactions (80 µl final volumes) Low FP. M III High FP.
Product Insert IMAP Screening Express Kit Product #8073 Quantity: 8000, 20 µl reactions (80 µl final volumes) Introduction About the IMAP Screening Express Kit The IMAP Screening Express Kit is used to
More informationab83355 ATP Assay Kit (Colorimetric/ Fluorometric)
Version 18 Last updated 2 January 2018 ab83355 ATP Assay Kit (Colorimetric/ Fluorometric) For the rapid, sensitive and accurate measurement of ATP in a variety of samples. This product is for research
More informationQuant-iT dsdna Broad-Range Assay Kit
Quant-iT dsdna Broad-Range Assay Kit Table 1. Contents and storage information. Material Amount Concentration Storage Stability Quant-iT dsdna BR reagent (Component A) λ DNA standards (Component C) Quant-iT
More informationFor the rapid, sensitive and accurate measurement of Ferrous and/or Ferric ions in various samples.
ab83366 Iron Assay Kit Instructions for Use For the rapid, sensitive and accurate measurement of Ferrous and/or Ferric ions in various samples. This product is for research use only and is not intended
More informationFor detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe.
ab112133 JC-10 Mitochondrial Membrane Potential Assay Kit Flow Cytometry Instructions for Use For detecting mitochondrial membrane potential changes in cells using our proprietary fluorescence probe. This
More informationFor the rapid, sensitive and accurate measurement of Citrate levels in various samples
ab83396 Citrate Assay Kit Instructions for Use For the rapid, sensitive and accurate measurement of Citrate levels in various samples This product is for research use only and is not intended for diagnostic
More information