製品の概要

  • 製品名
  • 製品の概要

    Avidin Biotin Blocking Kit ab64212 blocks signal from endogenous avidin, biotin and biotin-binding proteins in tissues when used with biotin-based IHC detection (eg. with ABC IHC detection kits).


    Find more kits and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.


     

  • アプリケーション
    適用あり: IHC-P, ICC/IF, IHC-Frmore details

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • バッファー
    Preservative: 0.08% Sodium azide
    Constituent: Avidin
  • 内容 15 ml
    Avidin Block 1 x 15ml
    Biotin Block 1 x 15ml
  • 研究分野
  • 関連性
    Some cells, and tissues such as kidney, liver and spleen, contain endogenous biotin. Using an avidin-biotin staining method may result in high, non-specific background staining. A significant reduction of this non-specific background can be obtained by pre-treatment of cells/tissues with avidin/biotin blocking reagents prior to the incubation of biotinylated antibody.

アプリケーション

Our Abpromise guarantee covers the use of ab64212 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

画像

  • Immunohistochemical analysis staining HTRA1 in mouse bone tissue sections. Tissue sections were dewaxed, rehydrated and treated with Endogenous Avidin/Biotin Blocking Kit (ab64212), 3% H2Oand normal swine serum. Tissue sections were then incubated with a polyclonal anti-HTRA1 antibody for 1 hour at 37°C. After PBS wash, tissue sections are incubated with biotinylated swine anti-rabbit IgG for 45 minutes at 37°C and further incubated for 30 minutes after washing. With Vectastatin. Sections were developed using DAB and counterstained using Harris modified hematoxylin. 

参考文献

This product has been referenced in:
  • Peng C  et al. Synchronous primary sigmoid colon cancer and primary thyroid cancer followed by a malignant tumor of the kidney: Case report of multiple primary cancer and review of the literature. Oncol Lett 17:2479-2484 (2019). Read more (PubMed: 30719116) »
  • Teissier T  et al. Knockout of receptor for advanced glycation end-products attenuates age-related renal lesions. Aging Cell 18:e12850 (2019). Read more (PubMed: 30794349) »
See all 6 Publications for this product

レビューと Q&A

1-4 of 4 Abreviews or Q&A

helpful kit

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used this kit to block the non-specific binding of avidin or biotinylated reagents with the endogenous biotin, also their receptors and the avidin binding sites in the tissue.
Its works good and is easy to use.
We used the kit as a ready to use-kit and incubated the samples first with the avidin block for 10minutes and after a washing step with the bioin block for 10minutes.

Abcam user community

Verified customer

投稿 Mar 12 2018

Answer


I can confirm that the Avidin Block and Biotin block provided with this kit come ready to use. They should not be diluted.

Once the sections are cut into sections and placed on a slide, the avidin and biotin blocking is done as part of the antibody staining procedure. You will need to pipette enough solution on to cover the tissue section and ensure it does not dry out during incubation. Typically this is usually 50 - 200 ul depending on the size of the section.

There is a protocol provided on the datasheet, see the link in the protocols tab.

Incubate tissue sections with Avidin Block for 10-15 minutes.
Wash 2 times with buffer.
Incubate tissue sections with Biotin Block for 10-15 minutes.
Wash 2 times with buffer
Apply Primary antibody to the tissue sections


I hope this will be helpful. If you have any further questions, please do not hesitate to contact me.

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Answer

Thank you for contacting us.

I would not recommend using the normal serum after biotin blocking for fear of introducing biotin to the samples. However if you would like, you may block with normal serum (5-10%) prior to biotin/avidin blocking. As you are not using a secondary you can use any normal serum at hand, donkey, goat, etc.

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

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Answer

Thank you for your reply.

In this case I would recommend that you quench endogenous biotin/avidin in your tissue prior to staining. We recommend use of our Endogenous Avidin/Biotin Blocking Kit (ab64212).

After blocking with this system, you may incubate your primary antibodies diluted in PSB or TBS.

Use of a mild detergent in your wash solution may also help to reduce background but isn't always neccessary.

More information about our Endogenous Avidin/Biotin Blocking Kit (ab64212) may be found at the location below:

https://www.abcam.com/Endogenous-Avidin-Biotin-Blocking-Kit-ab64212.html


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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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