Overview

  • Product name
    Anti-Biotin antibody (HRP)
    See all Biotin primary antibodies
  • Description
    Goat polyclonal to Biotin (HRP)
  • Host species
    Goat
  • Conjugation
    HRP
  • Tested applications
    Suitable for: Dot blot, ELISA, IHC-P, IHC-Fr, WBmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to Biotin conjugated to Keyhole Limpet Haemocyanin (KLH).

Properties

Applications

Our Abpromise guarantee covers the use of ab6651 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
Dot blot Use at an assay dependent concentration.
ELISA 1/140000.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB 1/3000 - 1/15000.

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

References

ab6651 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for contacting us.

While it is theoretically possible to amplify your signal by using a biotinylated secondary antibody with an anti-biotin tertiary antibody, it is not commonly done. Because streptavidin binds to biotin with a very high affinity, it is very commonly used to provide amplification in biotin systems in IHC or ELISA. For example, you could use streptavidin-HRP (ab64269) to detect the biotinylated secondary antibody. When using biotinylated secondary antibodies in WB, it is important to block with BSA instead of milk. Milk contains endogenous biotins that can cause very high background.

To increase the WB signal, it is more common to use a more sensitive ECL reagent or a longer exposure. Alternatively, you could try things like enriching your samples by subcellular fractionation(ab109719), loading more protein, using more primary antibody, or incubating overnight in the primary antibody. Some researchers also produce a slightly amplified signal by using a tertiary antibody. For example, you could use an unconjugated rabbit anti-mouse (ab97042)followed by an HRP conjugated anti-rabbit (ab97051).

I hope this helps, please let me know if you need any additional information or assistance.

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Answer

Thank you for contacting us. I see no reason why anti-biotin antibody ab6651 or HRP conjugated-streptavedin protein ab7403 would not recognise the biotin conjugated to your customers protein of interest following a boiling step. Biotinylated protein ladders such as this one: http://www.cellsignal.com/pdf/7727.pdf are denatured by heating and then detected by anti-biotin antibodies. Either product should be applicable to your customers Western blot proceedure. No special proceedure should be required to perform the Western blotting. A general proceedure can be found here: https://www.abcam.com/index.html?pageconfig=resource&rid=11375 I hope this information has been helpful. If you have any further questions please do not hesitate to ask.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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