Product nameHRP Conjugation Kit - Lightning-Link®
HRP Conjugation Kit ab102890 uses a simple and quick process to conjugate an antibody to HRP. It can also be used to conjugate other proteins or peptides.
To conjugate an antibody to HRP 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 HRP.
Amount and volume of antibody for conjugation to HRP
Kit size Recommended
amount of antibody1
amount of antibody
30 µg 3 x 10 µg 3 x 40 µg 3 x 10 µL 100 µg 100 µg 400 µg 100 µL 300 µg 3 x 100 µg 3 x 400 µg 3 x 100 µL 1 mg 1 mg 4 mg 1 mL 5 mg 5 x 1 mg 5 x 4 mg 5 x 1 mL
1 Recommended amount of antibody will give an HRP:antibody molar ratio of 4:1. Antibodies often also perform well at the maximum amount of antibody which is 1:1 molar ratio.
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.
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 azide2 PBS Potassium phosphate Sodium chloride HEPES Sucrose Sodium citrate EDTA Trehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.
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 100 µg 1 mg 30 µg 300 µg 5 mg HRP mix 1 x 100μg 1 x 1mg 3 x 10μg 3 x 100μg 5 x 1mg Modifier reagent 1 vial 1 vial 1 vial 1 vial 1 vial Quencher reagent 1 vial 1 vial 1 vial 1 vial 1 vial
- Horseradish peroxidase
Our Abpromise guarantee covers the use of ab102890 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.|
HRP conjugation kit ab102890 is used by Abcam extensively in ELISA development. It is used for development / manufacturing of Abcam's SimpleStep ELISA® kits, including the Frataxin ELISA kit ab176112 used to produce the image above.
Transformed B lymphocyte cells from Friedreich's Ataxia (FA) samples were compared to heterozygous carrier B lymphocyte cells (Carrier) and control B lymphocyte cells (Control). B lymphocyte cell extracts were analyzed across a 7-point titration (0.1-100 µg/mL) and frataxin levels were interpolated from the standard curve. Average interpolated values of Frataxin are plotted.
This product has been referenced in:
- Park JK et al. Evaluation of Preexisting Anti-Hemagglutinin Stalk Antibody as a Correlate of Protection in a Healthy Volunteer Challenge with Influenza A/H1N1pdm Virus. MBio 9:N/A (2018). Read more (PubMed: 29362240) ?
- Tan X et al. Rapid Mouse Follicle Stimulating Hormone Quantification and Estrus Cycle Analysis Using an Automated Microfluidic Chemiluminescent ELISA System. ACS Sens 3:2327-2334 (2018). Read more (PubMed: 30335974) ?