For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
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:
(check protocol booklet first)
- add modifier reagent to antibody and incubate for 3 hrs
- add quencher reagent and incubate for 30 mins
The conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification steps are required and 100% of the antibody is recovered for use.
Learn about buffer compatibility below; if your buffer is incompatible or antibody concentration is too low, please use our rapid antibody purification and concentration kits. Use our FAQs to learn more about the technology, or about conjugating other proteins and peptides to HRP.
Amount and volume of antibody for conjugation to HRP
(up to the following amounts)
|Kit size||Amount of antibody
at 4:1 molar ratio1
|Amount of antibody
at 1:1 molar ratio1
|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|
1 Optimal HRP:antibody molar ratio is 4:1. Antibodies often also perform well at a 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.
Sample Buffer Requirements
Compatible buffer constituents
(up to the concentrations shown, and pH 6.5-8.5):
|50mM Tris||0.1%/1% BSA*||0.1% gelatin|
|0.1% sodium azide**||PBS pH6.5-8.5||50% glycerol|
|0.15M sodium chloride||50mM HEPES||0.02M potassium phosphate|
|0.001% Tween||5% Trehalose||EDTA|
* 1% BSA gives lower quality conjugates.
** Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.
Incompatible buffer constituents
(in addition to any compatible constituents above the acceptable concentrations):
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.
|Components||100 µg||1 mg||30 µg||300 µg|
|HRP mix||1 x 100µg||1 x 1mg||3 x 10µg||3 x 100µg|
|Modifier reagent||1 vial||1 vial||1 vial||1 vial|
|Quencher reagent||1 vial||1 vial||1 vial||1 vial|
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.|
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"