Overview

  • Product name
    Biotinylation Kit / Biotin Conjugation Kit (Fast, Type B)
  • Product overview

    Biotinylation Kit / Biotin Conjugation Kit ab201796 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 B Biotinylation Kit / Biotin Conjugation Kit is optimized to produce conjugates for assays in which the biotinylated protein is captured by streptavidin immobilized on a surface (e.g., plates, nitrocellulose, magnetic beads etc).


    Use the Type A Biotinylation Kit / Biotin Conjugation Kit ab201795 to produce conjugates for assays in which a streptavidin-labeled detection reagent will be used.

  • Notes

    Amount and volume of antibody for conjugation to Biotin

     Kit size   Recommended 
    amount of antibody
    1 
    Maximum 
    amount of antibody
    Maximum antibody 
    volume2
    30 µg   3 x 10 µg  3 x 20 µg 3 x 10 µL
    300 µg   3 x 100 µg  3 x 200 µg 3 x 100 µL
    1 mg   1 mg 2 mg 1 mL

    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%/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 1% BSA gives lower quality conjugates, 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.

  • Tested applications
    Suitable for: Conjugationmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab201796 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Conjugation Use at an assay dependent concentration.

Images

  • This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.

Protocols

References

This product has been referenced in:
  • Lino Cardenas CL  et al. An HDAC9-MALAT1-BRG1 complex mediates smooth muscle dysfunction in thoracic aortic aneurysm. Nat Commun 9:1009 (2018). Read more (PubMed: 29520069) »
  • Islam F  et al. An electrochemical method for sensitive and rapid detection of FAM134B protein in colon cancer samples. Sci Rep 7:133 (2017). Read more (PubMed: 28273937) »
See all 2 Publications for this product

Customer reviews and Q&As

I have use the product Biotinylation Kit / Biotin Conjugation Kit (Fast, Type B) for biotinylation of Protein X. The product not only successfully biotinylated the protein, the protein can also be co-immunoprecipitated with streptavidin coated magnetic beads from cell lysate.

Abcam user community

Verified customer

Submitted Mar 13 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up