Cell Viability and Senescence Detection Kits

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2 Cell Viability and Senescence Detection Kits Cell viability and proliferation assays Growth factor/cytokine assays Cell culture condition optimization Cell number determination Multiwell and automation compatible PromoKine offers highly sensitive and convenient cell viability kits based on fluorometric (Resazurin and Calcein), colorimetric (WST-8, WST-1, XTT and MTT), and bioluminescent (ATP and ADP/ATP) detection. Additionally PromoKine also provides a kit for histochemical detection of senescent cells. Calcein AM R acetoxymethyl Esterase Viable cell Calcein Assay mechanism of the Fluorometric Cell Viability Kit II. Fluorometric Cell Viability Kit I (Resazurin) 500 assays 2,500 assays 10,000 assays PK-CA PK-CA PK-CA Viability Kit II (Calcein) 1,000 assays PK-CA Colorimetric Cell Viability Kit I (WST-8) Colorimetric Cell Viability Kit II (WST-1) Colorimetric Cell Viability Kit III (XTT) Colorimetric Cell Viability Kit IV (MTT) Bioluminescent Cell Viability Kit I (ATP) Bioluminescent Cell Viability Kit II (ADP/ATP) 500 assays PK-CA705-CK assays 2,500 assays PK-CA577-K PK-CA577-K ,000 assays PK-CA ,000 assays PK-CA assays 1,000 assays PK-CA577-K PK-CA577-K assays PK-CA577-K Senescence Detection Kit 250 assays PK-CA577-K

3 Live/Dead Cell Staining Kits Monitoring of viable and dead cells Cell counting by fluorescence microscopy Excitation and emission spectra are compatible with common excitation sources and filter sets The Live/Dead Cell Staining Kit II utilizes two fluorescent dyes: Calcein-AM and EthD-III. Calcein-AM penetrates the cell membrane of living cells, where it is hydrolyzed to cell membrane-impermeable, green-fluorescent Calcein (emission maximum at 515 nm). In contrast, Propidium Iodide only passes through the disrupted membranes of dead cells. In the nucleus it intercalates with the DNA and emits red light at 617 nm. Since both Calcein and EthD-III can be excited at 490 nm, a simultaneous monitoring of viable and dead cells is possible using a fluorescence microscope. Double-staining of viable (green) and dead (red) cells using the PromoKine Live/Dead Cell Staining Kit II. Live/Dead Cell Staining Kit II 300 assays PK-CA

4 Cytotoxicity Kits Cytotoxicity assays Chemosensitivity testing Toxicity screening Compatible with common plate readers LDH Cytotoxicity Kits PromoKine s easy-to-use colorimetric LDH Cytotoxicity Kits are based on the activity of lactate dehydrogenase (LDH), a stable cytoplasmic enzyme that is present in all cells. Damaged cells rapidly release this enzyme into the surrounding medium. LDH activity is determined by a coupled enzymatic reaction. It oxidizes lactate to pyruvate, triggering the conversion of the yellow tetrazolium salts INT or WST into red formazan dye. The increase in the amount of formazan can be analyzed spectrophotometrically. Lactate Formazan (red) LDH NAD + NADH + H + Catalyst Pyruvate WST-1 (yellow) Assay mechanism of the PromoKine LDH Cytotoxicity Kits. AK Cytotoxicity Kit The highly sensitive bioluminometric AK Cytotoxicity Kit is based on adenylate kinase (AK), a ubiquitous protein present in all cells that is rapidly released into the culture medium upon damage to the plasma membrane. The adenylate kinase release from the damaged cells catalyses the conversion of ADP to ATP. Luciferase then catalyses a light-producing reaction between the ATP, oxygen, and luciferin. This light can be measured using a luminometer or beta counter. LDH Cytotoxicity Kit II 500 assays PK-CA577-K AK Cytotoxicity Kit 500 assays PK-CA577-K

5 Cell-Based Reporter Assays Fast and convenient monitoring of reporter gene expression (Luciferase, GFP, ß-Galactosidase) Reliable determination of transfection efficiencies Luciferase Reporter Assay Kits The PromoKine ready-to-use Luciferase Reporter Assay Kits enable sensitive luciferase quantitation in eukaryotic cells, e.g., cells transfected with Firefly and/or Renilla luciferase reporter genes. High throughput kits are also available. GFP Reporter Assay Kit The kit has been designed for use in 96-well microplates. The amount of Green Fluorescent Protein (GFP) in cell and tissue samples is directly determined by comparing their fluorescence signals with a GFP standard. Beta-Galactosidase Kits The Beta-Galactosidase Kits have been designed to reliably detect and quantify ß-galactosidase activity in cells transfected with the LacZ reporter gene. ß-galactosidase can be detected microscopically (Staining Kit) or quantified using a spectrophotometer (Reporter Assay Kit). Luciferase Reporter Assay Kit I (Firefly) Luciferase Reporter Assay Kit II (Renilla) Luciferase Reporter Assay Kit III (Firefly & Renilla) Luciferase Reporter HTS Kit (Firefly) 150 assays 1,000 assays 150 assays 1,000 assays 100 assays 1,000 assays 1 ml 10 ml 100 ml 10 x 100 ml PK-CA PK-CA PK-CA PK-CA PK-CA PK-CA PK-CA PK-CA PK-CA PK-CA GFP Reporter Assay Kit 100 assays PK-CA577-K Beta-Galactosidase Reporter Assay Kit 500 assays PK-CA

6 Cell Staining Reagents Staining of viable and dead cells Detection of cellular structures Fluorescence microscopy/flow cytometry Thoroughly tested and highly sensitive PromoKine provides a wide range of high-quality fluorescent dyes and kits for staining different cell organelles and structures as well as for determining cellular processes: Nucleus Stains Mitochondria Stains ER Stains Viable Cell Stains Dead Cell Stains Cytoplasmic Membrane Stains Lipophilic Carbocyanine Dyes Phospholipid Membrane Probes Nerve Terminal Staining Kits Nerve Terminal Probes Membrane Potential Sensitive Dyes Cytosolic Tracers Primary human fibroblasts (NHDF, PromoCell) stained with Calcein and counterstained with DAPI.

7 Intracellular Indicators Signaling studies Neuroscience research High quality and sensitivity Investigation of signal transduction and apoptotic cascades The change of intracellular ion concentration is a crucial step that helps second messengers transmit information. Consequently, monitoring and control of intracellular ion concentration is a prerequisite for understanding second messengers and signal transduction in living cells. Promo- Kine provides the following high-quality intracellular indicators: Calcium indicators Near-membrane calcium indicators Zn indicators Heavy metal ion indicators ph indicators Cell Stress and Metabolism PromoKine provides sensitive and convenient kits for the following applications: GST activity measurement and quantitation of GSTtagged fusion proteins SOD assays Determination of the concentration of NO metabolites (nitrate/nitrite) HAT activity measurement Metabolic studies and analysis of mutagenic, carcinogenic, and toxic effects DNA damage detection and quantification Analysis of the cell redox state Lactate Determination ATP and ADP Quantification NAD/NADH and NADP/NADPH Quantification

8 Cell Signaling Quantitative determination of camp/cgmp levels Cell signaling and cell growth/differentiation studies ß-Secretase activity detection Alzheimer s disease drug development camp and cgmp Immunoassay Kits The PromoKine camp and cgmp competitive immunoassays have been developed for quantitative determination of camp and cgmp levels. They utilize polyclonal capture antibodies against camp or cgmp coated on a 96-well plate and HRP-conjugated camp or cgmp-hrp, which competes with the camp or cgmp from the sample. After HRP-substrate addition, the amount of camp-hrp or cgmp-hrp bound to the plate can easily be determined spectrophotometrically at 450 nm. ß -Secretase Fluorometric Assay Kit The PromoKine ß-Secretase Fluorometric Assay provides a convenient, non-radioactive system for detecting ß-secretase activity in mammalian samples. The kit provides active ß-secretase as positive control, ß-secretase inhibitor as negative control, optimized peptide substrate (conjugated to two reporter molecules), and buffers. camp Immunoassay Kit 100 assays PK-CA577-K cgmp Immunoassay Kit 100 assays PK-CA577-K Beta-Secretase Fluorometric Assay Kit Beta-Secretase Inhibitor I Beta-Secretase Inhibitor II 100 assays PK-CA577-K mg PK-CA mg PK-CA

9 Cell Fractionation and Protein Extraction Kits Cell Analysis Cellular protein extraction from mammalian cells Integral membrane protein extraction Preparation of highly enriched cellular fractions from mammalian cells PromoKine provides a wide range of cell fractionation and protein extraction kits. The isolated proteins and separated cellular fractions are suitable for many downstream applications such as 2-dimensional gel electrophoresis, protein immunoblotting, gel-shift assays, enzyme activity assays, reporter assays, and more. The following kits are available: Mammalian Whole Cell Protein Extraction Kit Membrane Protein Extraction Kit Nuclear/Cytosol Fractionation Kit Mitochondria/Cytosol Fractionation Kit Cytosol/Particulate Fractionation Kit Complete Cell Fractionation Kit Mammalian Whole Cell Protein Extraction Kit Membrane Protein Extraction Kit Nuclear/Cytosol Fractionation Kit Mitochondria/Cytosol Fractionation Kit Cytosol/Particulate Fractionation Kit Complete Cell Fractionation Kit 500 assays PK-CA577-K assays PK-CA577-K assays 100 assays 25 assays 100 assays PK-CA577-K PK-CA577-K PK-CA577-K PK-CA577-K assays PK-CA577-K assays PK-CA577-K

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