Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Overview
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Product name
Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® -
Product overview
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).
Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.
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Notes
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 antibody1Maximum
amount of antibodyMaximum antibody
volume23 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 > 2mg/ml or
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 If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.
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.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 1 mg 100 µg 3 x 100 µg 3 x 10 µg ab274076 - Biotin (Type A) Conjugation Mix 1 x 1mg 1 x 100µg 3 x 100µg 3 x 10µg ab273994 - Modifier reagent 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 -
Research areas
Images
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Conjugation - Biotin (Type A) Conjugation Kit (ab201795)
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
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Biotinylation Kit / Biotin Conjugation Kit (Type A) - Lightning-Link® labeling total proteins from lysate for a lectin microarray Image from Liu T et al., Front Physiol., 8:472. Fig 1.; doi: 10.3389/fphys.2017.00472. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/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.
