PE/Cy5® Conjugation Kit - Lightning-Link® (ab102893)
Overview
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Product name
PE/Cy5® Conjugation Kit - Lightning-Link® -
Product overview
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.
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Notes
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 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% 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 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 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 -
Research areas
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Alternative names
- Phycoerythrin
Images
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Conjugation - PE/Cy5® Conjugation Kit (ab102893)
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
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PE/Cy5® Conjugation Kit - Lightning-Link® labeling TcdA toxin for Flow cytometry Image from Olling A et al., PLoS One, 6(3):e17623. Fig 5.; doi: 10.1371/journal.pone.0017623. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/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.
