PE/Cy7® Conjugation Kit - Lightning-Link® (ab102903)
Overview
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Product name
PE/Cy7® Conjugation Kit - Lightning-Link® -
Product overview
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®.
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
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Notes
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® R-PE/Cy7 Labeling Kit. 762-0005 is the same as the 60 µg size. 762-0010 is the same as the 3 x 60 ug size. 762-0030 is the same as the 3 x 10 ug size. 762-0015 is the same as the 600 µg size.
Amount and volume of antibody for conjugation to PE/Cy7®.
Kit size Recommended
amount of antibodyMaximum antibody
volume13 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
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 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 If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.
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.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 60 µg 600 µg 3 x 10 µg 3 x 60 µg ab274106 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab274150 - PE/Cy7 mix 1 x 60µg 1 x 600µg 3 x 10µg 3 x 60µg ab274133 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl -
Research areas
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Alternative names
- Phycoerythrin
Images
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ab102903 PE-Cy7® conjugation kit used with a mouse anti-bovine CD45RO antibody Image from Oliveira BM et al., Scientific reports., 9 (1) 3413. Fig 1.; doi: 10.1038/s41598-019-39938-0. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.
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+
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PE/Cy7 KitThis illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.