Rabbit specific HRP/DAB (ABC) Detection IHC Kit (ab64261)

製品の概要

  • 製品名
    Rabbit specific HRP/DAB (ABC) Detection IHC Kit
    HRP/DAB detection kit キット 製品一覧
  • 製品の概要

    The reagents in this kit constitute a labeled streptavidin-biotin immunoenzymatic antigen detection system. This technique involves the sequential incubation of the specimen with an unconjugated primary antibody specific to the target antigen, a biotinylated secondary antibody which reacts with the primary antibody, enzyme-labeled streptavidin, and substrate-chromogen.

  • 特記事項

    DAB is a suspected carcinogen. Handle with care.

    Contains hydrogen peroxide

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

製品の特性

  • 保存方法
    Store at +4°C. Please refer to protocols.
  • 内容 15 ml
    50x DAB Chromogen 1 x 0.5ml
    Biotinylated goat anti-rabbit IgG(H+L) 1 x 15ml
    DAB substrate 1 x 15ml
    Hydrogen Peroxide Block 1 x 15ml
    Protein block 1 x 15ml
    Streptavidin Peroxidase 1 x 15ml
  • 研究分野
  • 関連性
    HRP/DAB detection kit is a immunoenzymatic antigen detection system for immunohistochemistry techniques.

アプリケーション

Our Abpromise guarantee covers the use of ab64261 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.
IHC-Fr Use at an assay dependent concentration.

画像

  • Immunohistochemical analysis staining TNF or IL6 in mouse tissue sections.  Samples are incubated in murine TNF (ab9739) or IL6 primary antibody. Bound primary antibodies were detected using IHC detection kit (ab64261). Immunohistochemistry was used to determine the effect of uridine on synovial expression of TNF and IL6. 

  • Immunohistochemical analysis of KLF4 and pVHL expression in colon cancer tissues. Sections from colon cancer and adjacent tissues were analyzed by immunostaining against KLF4 and pVHL. Staining without primary antibody served as negative control.

    Tissue sections were de-waxed with xylene and rehydrated through gradient ethanol into water. For antigen retrieval, sections were heated in citrate buffer (pH6.0) for 10 min at 95°C in a microwave oven. After cooling to room temperature, the sections were then digested with 0.05% trypsin for 10 min at 37°C. Endogenous peroxidase activity was quenched with 0.3% H2O2 in methanol for 30 min at room temperature. After PBS washes, nonspecific antibody binding was blocked by pre-incubating slides with 10% normal goat non-immune serum at 37° for 30 min. After blotting off the blocking serum, sections were incubated with primary antibody against LKF4 as well as primary antibody against VHL at 4° overnight. After PBS washes again, sections were incubated with biotinylated secondary antibody at 1:200 dilution for 30 min at room temperature. After incubating with Vectastain ABC reagent (Vector Laboratories, Inc., Burlingame, CA) for 30 min at room temperature, the sections were developed with Anti Rabbit HRP/DAB Detection Kit (ab64261). Sections were washed in running tap water and lightly counterstained with hematoxylin, followed by dehydration and coverslip mounting. Negative controls were obtained by omitting the primary antibody.

  • Human colon carcinoma fixed in 10% NBF for 24 hrs and stained with anti p53 antibody using Anti Rabbit HRP/DAB Detection Kit (ab64261).

参考文献

This product has been referenced in:
  • Savarin M  et al. Electrotransfer of plasmid DNA radiosensitizes B16F10 tumors through activation of immune response. Radiol Oncol 51:30-39 (2017). IHC-P ; Mouse . Read more (PubMed: 28265230) »
  • Uchibe K  et al. Genetic and pharmacological inhibition of retinoic acid receptor ? function promotes endochondral bone formation. J Orthop Res 35:1096-1105 (2017). Read more (PubMed: 27325507) »

See all 21 Publications for this product

レビューと Q&A

This issue may be result of sample preparation. It is oftentimes the case that the slides have not been inadequate deparaffinization which will lead to this type of staining. The link below outlines Abcam recommendations of deparaffinization: http:/...

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One drop of DAB chromogen is about 30 ul, and it is to be added to 1.5 ml of the substrate (50 drops). So the ratio is 1:50.

The mixture cannot be stored very long once you have made it. Ideally, you should mix it immediately before using it...

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I can confirm the Protein Block is a PBS solution with 0.5% Casein and 0.5% BSA.

I hope this information will be helpful for your customer.
Have a great day!

Thank you for getting in touch.

When using frozen sections, you can use a general frozen section staining protocol. I would amend the protocol booklet protocol as follows:

1. Fix the sections (for example 4% PFA for 10 minutes or ice ...

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It is very easy to use, get sharp images with it.

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit is very easy to use, just follow its instructions. It contains protein block, secondary antibody and substrate et al. in one box, basically everything you need for IHC except primary antibody. The box of the kit is of the right size, not too big not too small, so it is very easy to locate in the refrigerator. I wish the kit also contains Hematoxylin but it doesn't. The staining results are sharp and strong.
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Abcam user community

Verified customer

投稿 Nov 23 2016


The lab sent me the following explanations:

The protocol for ab64261 reflects the sequence we follow in house. Personally I have done the endogenous peroxidase blocking step before antigen retrieval, after antigen retrieval and after the...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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I am sorry that I was unavailable when you called but am glad to hear from you. We would not suggest using 5% BSA instead of the protein block included with the kit. These blocking solutions are formulated to give the best result with the kit. I...

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It does look like endothelium is stained, as expected, but yes, the background is high. Did you include a negative control, staining with just the secondary detection kit, and not the primary? If that is completely negative, and shows no background, th...

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Thank you for contacting us. You will need an antigen retrieval step. Customers have reported success with a few different protocols but I recommend a heat mediated retrieval using a 10mM citrate buffer, pH 6.0. This should be applied to the sections b...

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1-10 of 19 Abreviews or Q&A

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

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