Product nameBiotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link®
Biotinylation Kit / Biotin Conjugation Kit (ab201795) uses a simple and quick process to conjugate an antibody to Biotin. It can also be used to conjugate other proteins or peptides.
To conjugate an antibody to Biotin using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 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 Biotin.
The Type A Biotinylation Kit / Biotin Conjugation Kit has been optimized to produce conjugates for assays in which a streptavidin-labeled detection reagent will be used.
Use the Type B Biotinylation Kit / Biotin Conjugation Kit ab201796 for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (e.g., plates, nitrocellulose, magnetic beads etc).
Amount and volume of antibody for conjugation to Biotin
Kit size Recommended
amount of antibody1
amount of antibody
30 µg 3 x 10 µg 3 x 20 µg 3 x 10 µL 100 µg 1 x 100 µg 1 x 200 µg 1 x 100 µL 300 µg 3 x 100 µg 3 x 200 µg 3 x 100 µL 1 mg 1 mg 2 mg 1 mL
1 Using the maximum amount of antibody may result in less labelling per antibody.
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% 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 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 300 µg 1 mg 100 µg Biotin (Type A) Conjugation Mix 3 vials 3 vials 1 vial 1 vial Biotin (Type A) Modifier Reagent 1 vial 1 vial 1 vial 1 vial Biotin (Type A) Quencher Reagent 1 vial 1 vial 1 vial 1 vial
Our Abpromise guarantee covers the use of ab201795 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 concentration.|
This product has been referenced in:
- Martínez-López M et al. Microbiota Sensing by Mincle-Syk Axis in Dendritic Cells Regulates Interleukin-17 and -22 Production and Promotes Intestinal Barrier Integrity. Immunity 50:446-461.e9 (2019). Read more (PubMed: 30709742) »
- Giussani S et al. The Streptococcus agalactiae complement interfering protein combines multiple complement-inhibitory mechanisms by interacting with both C4 and C3 ligands. FASEB J 33:4448-4457 (2019). Read more (PubMed: 30566365) »