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

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (3)Q&A (13)References (62)

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Immunohistochemistry - ab64261
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/DAB Detection Kit (ab64261)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/DAB Detection Kit (ab64261)

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Overview

  • Product name

    Rabbit specific HRP/DAB (ABC) Detection IHC Kit
    See all HRP/DAB detection kit kits
  • Product overview

    Rabbit specific HRP/DAB (ABC) IHC Detection Kit ab64261 is a complete kit to detect a primary antibody raised in rabbit, using HRP-labeled-streptavidin and a biotinylated anti-rabbit secondary antibody.


    It is supplied with a full protocol and with peroxidase and protein blocking reagents, biotinylated secondary antibody, streptavidin-HRP, and DAB substrate.


    In the method:
    - a sample is incubated with an unconjugated primary antibody specific to the target antigen
    - a biotinylated secondary antibody then binds to the primary antibody
    - HRP-labeled streptavidin then binds to the secondary antibody
    - the HRP produces a brown colored substance at the site of primary antibody binding by reacting with DAB.

  • Notes

    For alternative streptavidin-biotin IHC detection kits see:
    - Anti-Mouse / anti-Rabbit HRP-DAB IHC kit ab64264
    - Anti-Mouse HRP-DAB IHC Kit ab64259

    For high sensitivity HRP micro-polymer IHC detection kits see:
    - Anti-Mouse / anti-Rabbit HRP/DAB Micro-polymer IHC Kit ab236466
    - Anti-Rabbit HRP-DAB Micro-polymer IHC Kit ab236469

    Alternatively, see HRP-AEC 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.

  • Tested applications

    Suitable for: IHC-P, IHC-Frmore details

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 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
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • IHC Kits
    • Kits/ Lysates/ Other
    • Kits
    • IHC tools
    • ABC
  • Relevance

    HRP/DAB detection kit is a immunoenzymatic antigen detection system for immunohistochemistry techniques.

Associated products

    Applications

    The Abpromise guarantee

    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.

    Application Abreviews Notes
    IHC-P
    Use at an assay dependent concentration.
    IHC-Fr
    Use at an assay dependent concentration.
    Notes
    IHC-P
    Use at an assay dependent concentration.
    IHC-Fr
    Use at an assay dependent concentration.

    Images

    • Immunohistochemistry - ab64261
      Immunohistochemistry - ab64261Image from Narendra S.C., PLoS One 10(10), Fig 5a. doi: 10.1371/journal.pone.0141863 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      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. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/DAB Detection Kit (ab64261)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Rabbit HRP/DAB Detection Kit (ab64261)This image is courtesy of an anonymous Abreview

      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.

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

    Protocols

    • Rabbit specific HRP/DAB detection kit
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (62)

    Publishing research using ab64261? Please let us know so that we can cite the reference in this datasheet.

    ab64261 has been referenced in 62 publications.

    • Jung YJ  et al. Cell reprogramming using extracellular vesicles from differentiating stem cells into white/beige adipocytes. Sci Adv 6:eaay6721 (2020). PubMed: 32232152
    • Shastri S  et al. Idebenone Protects against Acute Murine Colitis via Antioxidant and Anti-Inflammatory Mechanisms. Int J Mol Sci 21:N/A (2020). PubMed: 31940911
    • Kranjc Brezar S  et al. Electrotransfer of siRNA to Silence Enhanced Green Fluorescent Protein in Tumor Mediated by a High Intensity Pulsed Electromagnetic Field. Vaccines (Basel) 8:N/A (2020). PubMed: 32012775
    • Byun S  et al. Fasting-induced FGF21 signaling activates hepatic autophagy and lipid degradation via JMJD3 histone demethylase. Nat Commun 11:807 (2020). PubMed: 32042044
    • Negro G  et al. Molecular heterogeneity in breast carcinoma cells with increased invasive capacities. Radiol Oncol 54:103-118 (2020). PubMed: 32061169
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Ratings

    1-3 of 3 Abreviews

    Great product that gives best results each time.

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We are very happy using this kit. Every time without fail we get strong signals. perfectly drafted protocol for easy use. Needs optimization with time of incubation depending on the immunoreactivity and intensity of your primary antibody.

    Abcam user community

    Verified customer

    Submitted May 08 2018

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

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    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.

    Abcam user community

    Verified customer

    Submitted Nov 23 2016

    Research Associate

    Good Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    A review for the Rabbit specific HRP/DAB (ABC) Detection IHC Kit (ab64261)
    Immunohistochemical staining procedure
    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 DAB (Abcam, Cambridge, MA). 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.
    Figure legend
    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.

    Abcam user community

    Verified customer

    Submitted Jun 03 2014

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

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