Product namePE/Cy5® Conjugation Kit
PE/Cy5® Conjugation Kit ab102871 uses a simple and quick process to conjugate an antibody to PE/Cy5®. It can also be used to conjugate other proteins or peptides.
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 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®.
Amount and volume of antibody for conjugation to PE/Cy5®.
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 30 µg 600 µg 60 µg 180 µg Modifier reagent 1 vial 1 vial 1 vial 1 vial PE/Cy5 mix 3 vials 1 vial 1 vial 3 vials Quencher reagent 1 vial 1 vial 1 vial 1 vial
Our Abpromise guarantee covers the use of ab102893 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.|