Product namePE/Cy7® Conjugation Kit - Lightning-Link®
This product is manufactured by Expedeon, an Abcam company. Expedeon product code 762-0010 was previously called Lightning-Link® R-PE/Cy7 Labeling Kit 3 reactions each up to 60 µg and is the same as the 3 x 60 µg size of this product. Expedeon product code 762-0030 was previously called Lightning-Link® R-PE/Cy7 Labeling Kit 3 reactions each up to 10 µg and is the same as the 3 x 10 µg size of this product. Expedeon product code 762-0015 was previously called Lightning-Link® R-PE/Cy7 Labeling Kit 1 reaction up to 600 µg and is the same as the 600 µg size of this product. Expedeon product code 762-0005 was previously called Lightning-Link® R-PE/Cy7 Labeling Kit 1 reaction up to 60 µg and is the same as the 60 µg size of this product.
PE/Cy7® Conjugation Kit / PE/Cy7® Labeling Kit ab102903 uses a simple and quick process for PE/Cy7 labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.
To conjugate an antibody to PE/Cy7® using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The PE/Cy7® conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.
Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to PE/Cy7®.
Amount and volume of antibody for conjugation to PE/Cy7®.
Kit size Recommended
amount of antibody1
30 µg 3 x 10 µg 3 x 10 µL 60 µg 60 µg 60 µL 180 µg 3 x 60 µg 3 x 60 µL 600 µg 600 µg 600 µL
1 Kit is designed to give a 1:1 molar ratio of antibody to PE after conjugation.
2 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 5mg/ml or < 0.5 mg/ml should be diluted /concentrated.
The selling size of this product has been changed – it is now based on the amount of antibody that can be conjugated with the kit, not the amount of PE mix provided. The amount of antibody advised that can be used with the kit has also been updated to reflect what will give the best conjugation results. The quantity and formulation of reagents provided have not changed, if you have been previously using the kit successfully with a different amount of antibody, there is no need to change the way that you are using the kit.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Compatible buffer constituents
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.
50mM / 0.6% Tris1 0.1%/1% BSA2 50% glycerol 0.1% sodium azide PBS Potassium phosphate Sodium chloride HEPES Sucrose Sodium citrate EDTA Trehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 1% BSA gives lower quality conjugates, BSA can also interfere with the use of the conjugated antibody in tissue staining.
Incompatible buffer constituents
Thiomerosal Proclin Glycine Arginine Glutathione DTT
Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.
Tested applicationsSuitable for: Conjugationmore details
Storage instructionsStore at -20°C. Please refer to protocols.
Components 60 µg 600 µg 3 x 10 µg 3 x 60 µg 30 µg 180 µg Modifier reagent 1 vial 1 vial 1 vial 1 vial 1 vial 1 vial PE/Cy7 mix 1 x 100µg 1 x 1mg 3 x 10µg 3 x 100µg 3 x 10µg 3 x 100µg Quencher reagent 1 vial 1 vial 1 vial 1 vial 1 vial 1 vial
Our Abpromise guarantee covers the use of ab102903 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Conjugation||Use at an assay dependent dilution.|
Oliveira BM et al. used ab102903 PE-Cy7® conjugation kit with a mouse anti-bovine CD45RO antibody and ab102859 APC-Cy7® conjugation kit with a mouse anti-Bovine CD62L antibody. This enabled them to run their desired multicolor flow cytometry panel.
Data shows flow cytometry gating strategy used to define γδ T cells (TCRγδ+CD3+CD335−), CD4+ T cells (CD4+CD3+TCRγδ−CD335−), CD8+ T cells (CD8+CD3+TCRγδ−CD335−) and NK cells (CD335+CD3−) in the stromal vascular fraction (SVF) of mesenteric and subcutaneous bovine adipose tissue (MAT and SAT, respectively) and in peripheral blood leukocytes. Dead cells were excluded with Fixable Viability Dye (FVD), lymphocytes were gated based on SSC-A versus FSC-A and singlets were selected from the FSC-A versus FSC-H dot plot.
The flow cytometry gating strategy used to define CD45RO+ and CD62L+ T cell subpopulations is also shown in CD8+
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
This product has been referenced in:
- Oliveira BM et al. T cells in mesenteric and subcutaneous adipose tissue of Holstein-Friesian cows. Sci Rep 9:3413 (2019). Read more (PubMed: 30833655) »
- Xi H et al. IL-33 amplifies an innate immune response in the degenerating retina. J Exp Med 213:189-207 (2016). Read more (PubMed: 26755704) »