Product namePE/Cy5® Conjugation Kit - Lightning-Link®
PE/Cy5® Conjugation Kit / PE/Cy5® Labeing Kit ab102871 uses a simple and quick process for PE/Cy5 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/Cy5® using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The PE/Cy5 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/Cy5®.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® R-PE/Cy5 Labeling Kit. 760-0005 is the same as the 60 µg size. 760-0010 is the same as the 3 x 60 ug size. 760-0030 is the same as the 3 x 10 ug size. 760-0015 is the same as the 600 µg size.
Amount and volume of antibody for conjugation to PE/Cy5®.
Kit size Recommended
amount of antibody
3 x 10 µg 3 x 10 µg 3 x 10 µL 60 µg 1 x 60 µg 1 x 60 µL 3 x 60 µg 3 x 60 µg 3 x 60 µL 600 µg 1 x 600 µg 1 x 600 µL
1Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 1 mg/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% BSA 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%
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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 600 µg 60 µg 3 x 10 µg 3 x 60 µg ab274106 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab274144 - PE/Cy5 mix 1 x 600µg 1 x 60µg 3 x 10µg 3 x 60µg ab274133 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
Olling A et al. used ab102893 as part of examining the role of toxins in the pathogenicity of Clostridium difficile.
They used the kit to conjugate PE/Cy5® to TcdA toxin for use in flow cytometry.
Binding of fluorescent labeled TcdA-PE/Cy5 and TcdA1–1874-Atto488 to HT29 and CHO-C6 cells was investigated by FACS analysis. Right shift of the black curve illustrates toxin binding which was detected through fluorescence emission at 667 nm for TcdA and at 523 nm for TcdA1–1874, respectively. Due to different ratio of fluorophor and toxin, fluorescence intensity of TcdA-PE/Cy5 cannot directly be compared with TcdA1–1874-Atto488.
ab102893 has been referenced in 9 publications.
- Henrik Heiland D et al. Tumor-associated reactive astrocytes aid the evolution of immunosuppressive environment in glioblastoma. Nat Commun 10:2541 (2019). PubMed: 31186414
- He S et al. Clonal anergy of CD117+chB6+ B cell progenitors induced by avian leukosis virus subgroup J is associated with immunological tolerance. Retrovirology 16:1 (2019). PubMed: 30602379
- Sun C et al. Central role of IP3R2-mediated Ca2+ oscillation in self-renewal of liver cancer stem cells elucidated by high-signal ER sensor. Cell Death Dis 10:396 (2019). PubMed: 31113961
- Nielsen CM et al. Impaired NK Cell Responses to Pertussis and H1N1 Influenza Vaccine Antigens in Human Cytomegalovirus-Infected Individuals. J Immunol 194:4657-67 (2015). PubMed: 25855356
- White MJ et al. Differential activation of CD57-defined natural killer cell subsets during recall responses to vaccine antigens. Immunology 142:140-50 (2014). PubMed: 24843874
- Kakhlon O et al. Polyglucosan neurotoxicity caused by glycogen branching enzyme deficiency can be reversed by inhibition of glycogen synthase. J Neurochem 127:101-13 (2013). PubMed: 23607684
- Taichman RS et al. GAS6 receptor status is associated with dormancy and bone metastatic tumor formation. PLoS One 8:e61873 (2013). PubMed: 23637920
- Olling A et al. The repetitive oligopeptide sequences modulate cytopathic potency but are not crucial for cellular uptake of Clostridium difficile toxin A. PLoS One 6:e17623 (2011). PubMed: 21445253
- Ott S et al. Preparation of epoxy-based macroporous monolithic columns for the fast and efficient immunofiltration of Staphylococcus aureus. J Sep Sci 34:2181-92 (2011). PubMed: 21735547