Mouse specific HRP/DAB (ABC) Detection IHC Kit (ab64259)

Reviews (2)Q&A (17)References (33)
Immunohistochemistry - ab64259
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Mouse HRP/DAB Detection Kit (ab64259)

Overview

  • Product name

    Mouse specific HRP/DAB (ABC) Detection IHC Kit
    See all HRP/DAB detection kit kits

  • Product overview

    Mouse specific HRP/DAB (ABC) IHC Detection Kit ab64259 is a complete kit to detect a primary antibody raised in mouse, using HRP-labeled-streptavidin and a biotinylated anti-mouse 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-Rabbit HRP-DAB IHC Kit ab64261

    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.

    For staining mouse tissues with mouse antibodies, use mouse-on-mouse polymer kit ab127055.

    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-Pmore 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-mouse IgG(H+L) (ab64255) 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

  • 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 ab64259 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 dilution.
Notes
IHC-P
Use at an assay dependent dilution.

Images

  • Immunohistochemistry - ab64259
    Immunohistochemistry - ab64259Jovicic N et al., PLoS One 10(7). Fig 6D & E. DOI: 10.1371/journal.pone.0134089. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Immunohistochemical analysis staining SMA or CD68 in paraffin embedded mouse liver tissue sections Samples were incubated with anti-α- SMA (ab7817) or anti-CD68 (ab49777). A Mouse specific HRP/DAB (ABC) Detection IHC Kit (AB64259 was used as a stain and Mayer’s hematoxylin was used as a counterstain. Top panel of images staining CD68 and bottom panel staining α-SMA.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Mouse HRP/DAB Detection Kit (ab64259)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti Mouse HRP/DAB Detection Kit (ab64259)
    Human colon carcinoma stained with anti p53 antibody using Anti Mouse HRP/DAB Detection Kit (ab64259).

References (33)

Publishing research using ab64259? Please let us know so that we can cite the reference in this datasheet.

ab64259 has been referenced in 33 publications.

  • Wisgrill L  et al. Integrative transcriptome analysis deciphers mechanisms of nickel contact dermatitis. Allergy 76:804-815 (2021). PubMed: 32706929
  • Hou QQ  et al. TBC1D3 promotes neural progenitor proliferation by suppressing the histone methyltransferase G9a. Sci Adv 7:N/A (2021). PubMed: 33523893
  • Black EK  et al. The effect of dietary supplementation with high- or low-dose omega-3 fatty acid on inflammatory pathology after traumatic brain injury in rats. Transl Neurosci 12:76-82 (2021). PubMed: 33623714
  • Campelo T  et al. AMPAR-Dependent Synaptic Plasticity Initiates Cortical Remapping and Adaptive Behaviors during Sensory Experience. Cell Rep 32:108097 (2020). PubMed: 32877679
  • Pinto B  et al. Rescuing Over-activated Microglia Restores Cognitive Performance in Juvenile Animals of the Dp(16) Mouse Model of Down Syndrome. Neuron 108:887-904.e12 (2020). PubMed: 33027640
View all Publications for this product

Customer reviews and Q&As

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1-10 of 19 Abreviews or Q&A

DAB staining of human liver tissues

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
DAB staining of human liver tissues - sample was treated with Ms-ASGP-R Ab. (non-Abcam)

Dr. Armen Petrosyan

Verified customer

Submitted Aug 11 2021

Super easy to use, abtained nice image with it!

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
This IHC kit is very easy to use, I get very clear images the first time I use it. It basically has everything you need for IHC staining except mouse primary antibody. Using counter staining will make the image look sharper, please be aware that the kit does not have hematoxylin for nuclear counter staining.

Abcam user community

Verified customer

Submitted Nov 23 2016

Question

I've purchased Dab chromogen and stained only 14 slides, each slide has only one slice of
tissue ie each slide take only one drop of dab chromogen and then I
found that it ended up and could not complete the rest of the slides
of my own research.
Is
this is normal??? please solve the problem, I am most in need of it
and thank you.

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Answer

I understand you used all of the 0.5 ml of pure 50x DAB chromogen.
As stated in the protocol booklet, the DAB chromogen must be diluted 50 times before being applied on the samples. See step 8 on page 6 of the protocol booklet: "Add 30 μl (1 drop) DAB Chromogen to 1.5 ml (50 drops) of DAB Substrate, mix by swirling and apply to tissue".

To terminate the kit 64259, I would suggest to order the DAB substrate kit ab94665 (https://www.abcam.com/ab94665).

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Question

I have purchased the above product #ab64259 for use with IHC-P protocol and a mouse primary antibody. However, I would now like to use our own anti-mouse-HRP secondary antibody instead of the one in this kit, and then just use the DAB substrate/chromogen part.

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Answer


I have contacted the lab who have developed this kit. They have confirmed that you are free to use your own anti-mouse HRP secondary in place of the Biotinylated Goat anti-mouse antibody and Streptavidin Peroxidase supplied in the kit, as scientifically that would also work to detect the mouse primary antibody. However, the protocol recommendations stated on the ab64259 datasheet are optimized for the components included in the kit, so if you were to use slightly different components (different secondary antibody), you would have to be responsible for determining the optimal conditions.

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Question

Thank you for the reply. I have two more comments about this product:

1: The protocol calls for hematoxylin, but the kit does not supply any, nor does the protocol specify a concentration. Please either supply hematoxylin or state in the product description that the user will need to supply their own hematoxylin and a concentration to use.

2: The protocol calls for specifically measured amounts of DAB chromogen and DAB substrate, but these reagents are supplied in bottles with squeeze tops, making it impossible to pipet out the required amounts. In order to get these specific amounts, the squeeze caps have to be physically ripped off. Please remove the squeeze caps from these reagents.

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Answer

Thank you again for your feedback and comments.

We have now added hematoxylin to the catalog: ab128990.
https://www.abcam.com/Mayers-Hematoxylin-ab128990.html

As an apology to you for the inconvenience and an appreciation of your feedback,I'd be happy tooffer youa bottle of 60ml Hematoxylin free of charge.
Alternatively, a discount off DRAQ5™ (ab108410, https://www.abcam.com/DRAQ5™-50-µl-5mM-ab108410.html) whichcan be used for IHC staining can be offered together with the submission of an Abreview.

The other feedback you had sent will be looked into very soon as well.

I look forward to hear back from you.

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

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Question

Thank you for the reply. I have two more comments about this product:

1: The protocol calls for hematoxylin, but the kit does not supply any, nor does the protocol specify a concentration. Please either supply hematoxylin or state in the product description that the user will need to supply their own hematoxylin and a concentration to use.

2: The protocol calls for specifically measured amounts of DAB chromogen and DAB substrate, but these reagents are supplied in bottles with squeeze tops, making it impossible to pipet out the required amounts. In order to get these specific amounts, the squeeze caps have to be physically ripped off. Please remove the squeeze caps from these reagents.

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Answer

Thank you for your email.

I will pass on your suggestions to the according department in Abcam. Thank you so much for your feedback! We do appreciate this.

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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Question

buffer mentioned in kit:
Does it refers to PBS only or PBS + Tween 20?

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Answer

Thank you for contacting us.

Thewashing / rinsing buffer to be used with this kit isPBS/Tween 20.

We do carry this buffer in our catalog in 2 different sizes(125 ml and 1000 ml):

ab64028: https://www.abcam.com/20x-PBS-Buffer-with-Tween-20-ab64028.html
ab64247: https://www.abcam.com/20x-PBS-buffer-with-Tween-20-ab64247.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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Question

Would like to test antibody in IHC-P.

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Answer

DISCOUNT CODE: XXXXXXXXXXX
Expiration date: 21-09-2012

I am very pleased to hear you would like to accept our offer and test ab89566 in [IHC-P]. This code will give you: 1 free [PRIMARY ANTIBODY/PROTEIN] OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for [IHC-P] and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question

Bonjour,

Tout d'abord merci pour votre aide.

J'ai effectué les tests avec votre kit de révélation pour la NGF et la TRkA avec les paramètres suivant :

- 3*5min a 600w dans un tampon à pH9

- 30 min de peroxydase

- 1h de serum block

- 30 min anticorps primaire

- 15 min biotine

- 15 min streptadivine

- DAB, Hematoxyline etc.

Nous avons tester sur le cerveau, sur la peau et sur de la muqueuse gingival. Nous avons un marquage mais nous ne sommes pas sur du résultat car dans la muqueuse l'épithélium est intégralement marqué. Est ce normal ?
De plus notre contrôle négatif, qui vient de chez vous,(ab47472) à aussi un léger font.

Les conditions si-dessus sont celle qui on donné le meilleur résultat. Pouvons nous vous envoyer les coupe afin que vous nous donniez votre avis ?

Merci d'avance pour votre aide.

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Answer

Merci pour votre émail.

Je suis désolée que les résultats ne sont pas claires.

Je vais volontiers investiguer ces résultats et essayer de donner des conseils pour améliorer les images. Si un de nos produits serait défectueux, nous le remplacerons en accord avec notre garantie.

Pour mieux comprendre la problème, est-ce que vous pouvez s'il vous plaît m'envoyer les images annotées de ces résultats? C'est plus pratique si vous pouvez nous envoyer les images, et non les slides.

Aussi, pourriez vous s'il vous plait dire quels anticorps exactement vous avez utilisés pour les marquages avec quels kits? C'était le ab6199 ou le ab3125?

Est-ce que vous avez utilisé des coupes paraffine de souris? Quel sérum est-ce que vous avez utilisé pour le blocage? Est-ce que vous avez aussi utilisé le protéine block du kit?

Je me réjouis de votre réponse.

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Question

Thank you.

I will follow your links.

I asked about prices because I might have the order via relatives in Canada or China.

Many thanks.

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Answer

Thank you for your reply.

If you change the country at the top right hand site of the Abcam website (https://www.abcam.com), you can see the country specific prices and also (in case of China) the distributor details.

Please be aware that we might not be able to ship to residential addresses.

I wish you good luck with your research.

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1-10 of 19 Abreviews or Q&A

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