Overview

  • Product name

  • Description

    Rabbit polyclonal to Biotin
  • Host species

    Rabbit
  • Specificity

    Reacts with free biotin and biotinylated antibody and proteins
  • Tested applications

    Suitable for: WB, Competitive ELISA, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Full length protein conjugated to KLH

Properties

Applications

Our Abpromise guarantee covers the use of ab53494 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
WB Use at an assay dependent dilution. Predicted molecular weight: 1 kDa.
Competitive ELISA Use at an assay dependent dilution. (sensitivity: 0.01 ng/mL).
IP Use at an assay dependent dilution.
ICC/IF Use at an assay dependent dilution.

Target

  • Relevance

    Biotin is a water soluble vitamin, generally classified as a B complex vitamin, also called vitamin B4. After the initial discovery of biotin, nearly forty years of research were required to establish it as a vitamin. Biotin is required by all organisms but can only be synthesized by bacteria, yeasts, molds, algae, and some plant species. Biotin is required as prosthetic group of enzymes involved in incorporation of carbon dioxide into organic compounds. Biotin has a MW of 244 Da.
  • Alternative names

    • Vitamin B4 antibody

Images

  • RWP1 cells were lysed with Polysomal Lysis
    Buffer and diluted in binding buffer (20 mM Hepes
    pH 7.6, 150 mM KCl, 3 mM MgCl2, 10% glycerol, 3 mg/ml BSA, 0.2 Triton X-100, 20 µg/ml poly dI-dC, 1 mM DDT, plus protease and RNase inhibitors). Pre-clearing was performed by incubating with protein G-agarose blocked with salmon sperm (1 mg/mL) and mouse IgG (10 µg/mL). After pre-clearing, samples (500 µg) were incubated 4 hours in binding buffer with the DNA-biotinylated probes (2 µg) and antibody against biotin (20 µg/mL) in a final volume of 1 mL. Samples were pulled-down with protein G-agarose, washed with binding buffer supplemented with 0.1%
    Tween-20 and analyzed by Western Blot for Suz12.

    See Abreview

References

This product has been referenced in:

  • Galbiati A & d'Adda di Fagagna F DNA Damage In Situ Ligation Followed by Proximity Ligation Assay (DI-PLA). Methods Mol Biol 1896:11-20 (2019). Read more (PubMed: 30474835) »
  • Mukherjee A  et al. Understanding How Wnt Influences Destruction Complex Activity and ß-Catenin Dynamics. iScience 6:13-21 (2018). Read more (PubMed: 30240607) »
See all 17 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Human Cell lysate - whole cell (RWP1)
Specification
RWP1
Total protein in input
500 µg

Abcam user community

Verified customer

Submitted Feb 14 2014

Answer

Thank you for contacting us.

The optimal dilutions or concentrations has to be determined by end user. We can recommend using the antibody at starting dilution 10ug/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

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