Product nameBiotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link®
Biotinylation Kit / Biotin Conjugation Kit (Type A) (ab201795) uses a simple and quick process for biotinylation / biotin labeling 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 Biotin using this kit:
- add modifier to antibody and incubate for 15 mins
- add quencher and incubate for 5 mins
The biotin 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).
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Rapid Biotin Type A Labeling Kit. 370-0005 is the same as the 100 µg size. 370-0010 is the same as the 3 x 100 ug size. 370-0030 is the same as the 3 x 10 ug size. 370-0015 is the same as the 1 mg size.
Amount and volume of antibody for conjugation to Biotin
Kit size Recommended
amount of antibody1
amount of antibody
3 x 10 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 3 x 100 µg 3 x 100 µg 3 x 200 µg 3 x 100 µL 1 x 1 mg 1 x 1 mg 1 x 2 mg 1 x 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 1 mg 100 µg 3 x 100 µg 3 x 10 µg 30 µg ab274076 - Biotin (Type A) Conjugation Mix 1 x 1mg 1 x 100µg 3 x 100µg 3 x 10µg 3 x 10µg ab273994 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab273995 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
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 illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
Liu T et al. used ab201795 as part of examining glycobiomarker in hepatocellular carcinoma metastasis.
They used the kit to conjugate biotin to total proteins from lysate for use in a lectin microarray.
Lectin microarray analysis of glycoforms. (A) The lectin microarray contains 50 lectin spots with different binding specificities. (B) Scan image of the lectin microarray incubated with biotinylated proteins and Cy5 labeled streptavidin. Typical glycan profiles of HCC with metastasis were shown on the right and HCC with non-metastasis on the left. (C) Hierarchical clustering of positive lectin binding spots (S/B≥2). Each row represented a single lectin, S/B values were shown by the color scale: red represents a lectin with high S/B value while green represents a lectin with low S/B value.
ab201795 has been referenced in 9 publications.
- 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). 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). PubMed: 30566365
- Karbanowicz TP et al. Comparison of Protein Gut Samples from Rhipicephalus spp. Using a Crude and an Innovative Preparation Method for Proteome Analysis. Vet Sci 5:N/A (2018). PubMed: 29538322
- Cornec D et al. Identification and phenotyping of circulating autoreactive proteinase 3-specific B cells in patients with PR3-ANCA associated vasculitis and healthy controls. J Autoimmun 84:122-131 (2017). PubMed: 28870527
- Drachsler M et al. CD95 maintains stem cell-like and non-classical EMT programs in primary human glioblastoma cells. Cell Death Dis 7:e2209 (2016). PubMed: 27124583
- Zhang A et al. Development and application of a fluorescence protein microarray for detecting serum alpha-fetoprotein in patients with hepatocellular carcinoma. J Int Med Res 44:1414-1423 (2016). PubMed: 27885040
- Jones SI et al. Direct detection of transcription factors in cotyledons during seedling development using sensitive silicon-substrate photonic crystal protein arrays. Plant Physiol 167:639-49 (2015). PubMed: 25635113
- Zhao Y et al. Protein microarray with horseradish peroxidase chemiluminescence for quantification of serum a-fetoprotein. J Int Med Res 43:639-47 (2015). PubMed: 26198141
- Hansen U et al. WARP interacts with collagen VI-containing microfibrils in the pericellular matrix of human chondrocytes. PLoS One 7:e52793 (2012). PubMed: 23300779