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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
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|
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%/1% BSA2||50% glycerol|
|0.1% sodium azide3||PBS||Potassium phosphate|
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.
3 Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.
Incompatible buffer constituents
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.
|组件||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"