Lightning-Link The world s easiest to use antibody labeling kits

Transcription

1 Lightning-Link The world s easiest to use antibody labeling kits InnovaCoat GOLD A revolution in nano-gold conjugation Thunder-Link The easiest path to oligo conjugation

2 Lightning-Link The world s easiest to use antibody labeling kits Wide range of labels Label from 10μg up to 1g of antibody or protein 30 seconds hands-on time 100% antibody recovery as no separation step is required 300+ references citing Lightning-Link Antibody Labeling Process Antibodies Proteins Peptides Add the Antibody Lightning-Link Label Fast Labeled Antibody Easy to use 1 Reliable

3 Lightning-Link Fluorescent Labeling Kits Lightning-Link Lightning-Link Rapid Colors Maximal Absorbance (nm) Maximal Emission (nm) DyLight 350 (320-xxxx) AMCA (712-xxxx) Atto 390 (349-xxxx) DyLight 405 (321-xxxx) PerCP (718-xxxx) PerCP/Cy5.5 (763-xxxx) PE/Cy5 (760-xxxx) DyLight 488 (322-xxxx) Fluorescein (707-xxxx) (310-xxxx) Atto 488 (733-xxxx) (350-xxxx) R-Phycoerythrin (RPE) (703-xxxx) PE/Texas Red (767-xxxx) PE/Atto 594 (768-xxxx) PE/Cy5.5 (761-xxxx) PE/Cy7 (762-xxxx) Rhodamine (710-xxxx) (311-xxxx) B-Phycoerythrin (BPE) (716-xxxx) Cy3 (786-xxxx) (340-xxxx) DyLight 550 (323-xxxx) Atto 565 (351-xxxx) FluoProbes 594 (790-xxxx) DyLight 594 (324-xxxx) Texas Red (714-xxxx) Atto 633 (353-xxxx) APC/Cy5.5 (764-xxxx) APC/Cy7 (765-xxxx) DyLight 633 (325-xxxx) Cy5 (781-xxxx) (342-xxxx) Allophycocyanin (APC) (705-xxxx) DyLight 650 (326-xxxx) FluoProbes 647H (791-xxxx) (362-xxxx) Cy5.5 (782-xxxx) DyLight 680 (327-xxxx) Atto 700 (751-xxxx) DyLight 755 (328-xxxx) DyLight 800 (329-xxxx) *The kits come in two formats - Lightning-Link and Lightning-Link Rapid, with incubation times of 3 hours and 15 minutes respectively. 2

4 Flow Cytometry Data of Lightning-Link Conjugated and Commercially Conjugated Antibodies A mouse monoclonal antibody, clone RPA-T4, specific for human CD4 was purchased from a commercial source in both unconjugated and RPE conjugated formats. The unconjugated antibody was linked to RPE using a Lightning-Link kit, and the two conjugates were compared in flow cytometry, staining human peripheral blood lymphocytes as shown above. Lightning-Link conjugated antibody is shown on the left hand side, and commercially conjugated antibody on the right hand side The blue curve shows unstained cells. A mouse monoclonal antibody, clone HCD54, specific for human CD54 was purchased from a commercial source in both unconjugated and RPE conjugated formats. The unconjugated antibody was linked to RPE using a Lightning-Link kit, and the two conjugates were compared in flow cytometry, staining human peripheral blood lymphocytes as shown above. Lightning-Link conjugated antibody is shown on the left hand side, and commercially conjugated antibody on the right hand side. The blue curve shows unstained cells. A mouse monoclonal antibody, clone RPA-T4, specific for human CD4 was purchased from a commercial source in both unconjugated and FITC conjugated formats. The unconjugated antibody was linked to fluorescein using a Lightning-Link kit, and the two conjugates were compared in flow cytometry, staining human peripheral blood lymphocytes as shown above. Lightning-Link conjugated antibody is shown on the left hand side, and commercially conjugated antibody on the right hand side. The blue curve shows unstained cells. 3

5 Lightning-Link Labeling Kits Lightning-Link Enzymes Horseradish Peroxidase (HRP) (701-xxxx) Alkaline Phosphatase (AP) (702-xxxx) Glucose Oxidase (706-xxxx) Lightning-Link Biotin/Streptavidin Avidin (717-xxxx) Biotin (704-xxxx) Streptavidin (708-xxxx) Western-Blotting-Enhanced Sensitivity Using Lightning-Link Conjugates Product Directly Labeled Antibodies Indirectly Labeled Antibodies Primary antibody Goat Anti-NQ01 Goat Anti-NQ01 Target quinone reductase 1 quinone reductase 1 Sample lysate human kidney human kidney Primary antibody working concentration µg/ml 0.1 µg/ml Secondary antibody used no (direct conjugation) yes Exposure time (min) Primary antibody source Everest Biotech, Cat no: EB05370 Primary antibody directly conjugated to HRP using Lightning-Link shows significantly enhanced sensitivity in Western blotting in comparison with traditional indirect technique. Enhanced ELISA Sensitivity Using Lightning-Link Conjugates LL kit generated antibody LL kit generated antibody with no antigen Commercially available antibody Commercially available antibody with no antigen 4

6 InnovaCoat GOLD A revolution in nano-gold conjugation Proprietary surface coat Ultra-stable Enhanced assay sensitivity Covalently link antibodies and analytes Simple conjugation process Different surface chemistries Diverse applications including lateral flow, flow cytometry, and electron microscopy InnovaCoat GOLD = Increased Sensitivity A comparison of antibody conjugation using InnovaCoat GOLD to passive conjugation techniques with naked gold, showing enhanced signal intensity. 40nm gold particles were labelled with anti-iga antibody, and used to measure IgA concentration in a lateral flow inhibition assay, with IgA bound to a lateral flow strip. The data shows the enhanced signal seen using InnovaCoat in comparison to passive gold conjugation. Increasing amounts of IgA in the buffer matrix result in a reduction in signal that can be used to measure IgA concentration. The enhanced curve seen with InnovaCoat GOLD provides a more sensitive and accurate measure of the IgA concentration. 5

7 Nano-Gold Particles Add the Antibody Labeled Nano-Gold Product Nanoparticle Size Target Chemistry Lateral Flow Electron Microscopy Flow Cytometry InnovaCoat GOLD 20nm Amine InnovaCoat GOLD 40nm Amine InnovaCoat GOLD - Maleimide 40nm sulfhydryl groups InnovaCoat GOLD - Hydrazide 40nm aldehydes* InnovaCoat GOLD Biotin 40nm Not applicable InnovaCoat GOLD Streptavidin 40nm Not applicable InnovaCoat GOLD 80nm Amine 4 4 (* kit converts Fc antibody carbohydrates to aldehydes) Options for labeling gold nanoparticles using InnovaCoat s various surface chemistries Standard Labeling InnovaCoat GOLD InnovaCoat GOLD - Hydrazide InnovaCoat GOLD - Maleimide 6

8 Thunder-Link The easiest path to oligo conjugation Thunder-Link kits for generating oligonucleotide conjugates End user chooses oligo sequence and antibody Applicable to all isotypes and species derived antibodies Oligo sizes (HPLC purified) between bases Amine group positioned at the 5 or 3 Positive control included All necessary components for a successful conjugation supplied Ideal for oligo-linked immunosorbent assays Optional step for free oligo removal post conjugation is included in the kit Advantages of Oligo-Linked Immunosorbent Assays in comparison with traditional ELISA - Enhanced Sensitivity + Small Amounts of Starting Material ELISA Oligo-Linked Immunosorbent Assay* Detection Limits µg to pg pg to fg Amount of starting material µl pl *Combines the specificity of antibodies with the amplification power of PCR 7

9 Product Conjugate to Modification required Thunder-Link (420-xxxx) Antibody of your choice Amine Thunder-Link GOLD (430-xxxx) 40nm Gold nanoparticles Sulfhydryl Simple Conjugation Process Thunder-Link process 8

10 Accessory Products Antibody Purification Kits: A complete range of simple to use kits that enable you to purify and concentrate any antibody that does not fit our Lightning-Link requirements Activated Labels: An alternative to the Lightning-Link chemistry for those who need to use sulfhydryl chemistry provided in ultra-stable freeze-dried format Imm-Link Kits: Conjugate your hapten to BSA, KLH or Ovalbumin. Available in 3 different chemistries, perfect for preparing antigens for immunisation HRP LifeXtend Stabilizer: Prolong the shelf life of your HRP conjugates HRP conjugated goat anti-rabbit IgG (1mg/ml) was diluted 1/10,000 in each of four stabilizers and incubated for 12 days at 37 C. An undiluted control sample was incubated at 4 C for the same period and diluted at 1/10,000 prior to ELISA testing. Conjugates were incubated on a rabbit IgG-coated plate for 1 hour at room temperature, followed by washing and determination of HRP activity using ABTS substrate. As can be seen above, even some of the best known stabilisers perform either poorly or only modestly in standard accelerated stability trials at 37 C. LifeXtend HRP stabiliser/ diluent offers the greatest protection and can substantially extend the shelf-life of HRP conjugates stored at 4 C or at room temperature. 9

11 Custom Services Conjugation: Take advantage of our expertise for your specialist conjugation requirements Optimisation: Select the best possible formulation to get the optimal conjugate of YOUR antibody for YOUR application (applicable to all 3 main product lines) Antibody Fragmentation: Our team of experts can generate Fab or F(ab) 2 fragments from your antibody for your specialist assay needs Bespoke Kits: We are able to provide larger kit sizes, and unique formulations based on your particular requirements and even apply our Lightning-Link technology to your own label Inventory Management: We offer batch validation and reservation according to your requirements URS is a member of Registrar of Standards (Holdings) Ltd. In recognition of our continuing commitment to quality, Innova Biosciences has been awarded with ISO 9001:2008. The customer can therefore be confident that they will receive high quality products and related services from Innova Biosciences that consistently meet customer and regulatory requirements. 10

12 Testimonials Lightning-Link Alkaline Phosphatase kit is a simple, easy-to-use kit, to biochemically label protein of interest, in our case, anti-hiv envelope antibodies, for ELISA. Labeling antibody with this kit does not affect the function and avidity of the antibody. One advantage in using this kit is that there is no need to remove excess alkaline phosphatase from the reaction mixture at the end of the incubation. The omission of this step does not affect our ELISA analysis. Tommy Tong, Post-Doctoral Fellow Having made our own passively adsorbed antibody-coated gold colloid for some years then more recently moving to a high OD, no-spin method, we were a little reluctant to try out another method quite so soon. However the extreme simplicity and rapidity of the InnovaCoat GOLD covalent conjugation, combined with the excellent functionality and consistency of the final product greatly helped us make the change. We now use less colloid per test in a demanding, room temperature stored, liquid formulation and are finding the development of new rapid tests to be significantly easier. Phil Goodwin, Chief Executive at Bio-Check (UK) Limited We have been working with Innova for about a year now and have had nothing but fantastic experiences with both their sales staff as well as their R&D staff. From the timely generation of quotes and custom project pricing, to the extremely helpful discussions with R&D, we have been duly impressed. Everyone on the Innova team handles themselves with the utmost professionalism and courtesy, and are experts in field. Regarding custom antibody conjugates, we were initially impressed with the quality of the conjugates we ordered, and are constantly being re-impressed since the quality of subsequent conjugates keeps improving! Again, the Innova team s expertise in the challenging field of antibody conjugation allows them to tackle any project we toss to them with no hesitation and generate a top-notch product to boot. Overall, we are thoroughly pleased with Innova s work and staff. We would wholeheartedly recommend them to anyone who needs excellent custom labeling and conjugation work you will not be disappointed! Lou Casta PhD, Project Leader at Genisphere, LLC. Innova Biosciences products are sold for research purposes only, and our terms and conditions of sale include a limited use license to our Intellectual Property for internal research applications. Commercial use, such as use within manufacturing, re-sale to third parties, or incorporation into kits, requires a separate written agreement, conferring relevant additional rights, with Innova Biosciences. Lightning-Link, InnovaCoat and Thunder-Link are registered trademarks of Innova Biosciences Cy and CyDye are trademarks of GE Healthcare DyLight is a registered trademark of Thermo Fisher Scientific Inc. and its subsidiaries FluoProbes is a registered trademark of interchim Texas Red is a registered trademark of Life Technologies Corporation