Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC kit (ab64264)

Overview

  • Product name
    Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC kit
    See all HRP/DAB detection kit kits
  • Product overview

    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.

  • Notes

    DAB is a suspected carcinogen. Handle with care.

    Contains hydrogen peroxide.

  • Tested applications
    Suitable for: IHC-Pmore details

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 15 ml 15 ml 15 ml
    50x DAB Chromogen 1 x 0.5ml 1 x 0.5ml 1 x 0.5ml
    Biotinylated Goat Anti-Polyvalent 1 x 15ml 1 x 15ml 1 x 15ml
    DAB substrate 1 x 15ml 1 x 15ml 1 x 15ml
    Hydrogen Peroxide Block 1 x 15ml 1 x 15ml 1 x 15ml
    Protein block 1 x 15ml 1 x 15ml 1 x 15ml
    Streptavidin Peroxidase 1 x 15ml 1 x 15ml 1 x 15ml
  • Research areas
  • Relevance
    HRP/DAB detection kit is a immunoenzymatic antigen detection system for immunohistochemistry techniques.

Applications

Our Abpromise guarantee covers the use of ab64264 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.

Images

  • Immunohistochemical analysis staining CD163 in mouse soleus muscle section. Samples were incubated in rabbit CD163 antibody at 1:450 dilution and visualized using mouse and rabbit specific HRP/DAB detection IHC Kit (ab64264).

  • Staining of human estrogen receptor in breast carcinoma tissue using ab16660 - Rabbit monoclonal [SP1] to Estrogen Receptor, and ab64246 - Mouse & Rabbit HRP/DAB Detection Kit.

References

This product has been referenced in:
  • Tong WY  et al. Delivery of siRNA in vitro and in vivo using PEI-capped porous silicon nanoparticles to silence MRP1 and inhibit proliferation in glioblastoma. J Nanobiotechnology 16:38 (2018). Read more (PubMed: 29653579) »
  • Nordquist E  et al. Postnatal and Adult Aortic Heart Valves Have Distinctive Transcriptional Profiles Associated With Valve Tissue Growth and Maintenance Respectively. Front Cardiovasc Med 5:30 (2018). IHC-P . Read more (PubMed: 29740591) »
See all 29 Publications for this product

Customer reviews and Q&As

1-10 of 10 Q&A

Answer


The pretreatment note in the protocol of the IHC kit refers to the antigen retrieval step that is required for most antibody to achieve a successful IHC-P staining.
For both of these antibodies (ab14440 and ab52894) I can recommend to use Sodium Citrate Buffer pH 6.0.
Please find more information on antigen retrieval here:
https://www.abcam.com/index.html?pageconfig=resource&rid=11488
We offer 10 x Sodium Citrate Buffer pH 6.0 for this purpose:
ab64214 (10x Citrate Buffer pH 6.0) https://www.abcam.com/index.html?datasheet=64214 https://www.abcam.com/index.html?datasheet=64214.
Please note that this step often requires intense optimisation and I can recommend to try a time curve to find the best antigen protocol for your samples and antibodies.
I wish you good luck with your research.

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Answer

This is confusing. ab113316 is a kit not antibody, customer would not be requiring a DAB kit for this.

For ab101854, ab80436 will be a good product.

Thanks!

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Answer

Thank you for contacting us.

The kits ab80436 and ab64264, both will be OK to use with ab10148 only because it is rabbit antibody, you can select any one of them. Please note that these kits cannot be used with ab75714 because it is a chicken polyclonal antibody - you would require a anti chicken HRP conjugated antibody.

There are two possibilities one you change ab75714 with mouse or rabbit as host, second buy the HRP conjugated anti rabbit and anti chicken secondary antibodies including DAB reagents.

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


Yes, Mouse and Rabbit Specific HRP/DAB (ABC) detection IHC kit (ab64264) and AEC Substrate System (Ready to Use) (ab64252) can be used with rat sections.

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Answer

Thank you for your enquiry.

The list of components of this kit can be found on the online datasheet:
https://www.abcam.com/index.html?datasheet=64264 (or use the following: https://www.abcam.com/index.html?datasheet=64264).

50x DAB Chromogen 1 x 0.5ml
Biotinylated goat anti-mouse & rabbit IgG(H+L) in PBS with stabilizers 1 x 15ml
DAB substrate 1 x 15ml
Hydrogen Peroxide Block 1 x 15ml
Protein Block (contains casein) 1 x 15ml
Streptavidin Peroxidase 1 x 15ml

It contains blocking solutions and detection reagents only. The customer will therefore require in addition to this an antigen retrieval solution, PBS, antibody diluentand primary antibody and possibly further blocking solution. I can suggest they review their IHC protocol to ensure they have all the items they require.

The following page from our website lists ourproducts for IHC which I hope will be helpful:
https://www.abcam.com/index.html?pageconfig=resource&rid=12853

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.

The protocol mentioned described on the datasheet refers to an Abreview from a previous user of this antibody. Different protocols may also work.

If you are using ab64264 kit, there is no need to use an extra blocking agent, as the kit already contains a protein block buffer to avoid unspecific bindings.

Regarding the antibody dilution, you can either use the ready to use antibody diluent or the general recommended diluent buffer (1% BSA in PBST).

You can use our online protocols as a general guidance:

https://www.abcam.com/index.html?pageconfig=resource&rid=11384

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 enquiry which has been forwarded to the Scientific support team.

I am sorry, it is regrettably not possible toprovide this particular component separately.

If you have any further questions, please do not hesitate to contact me.

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Answer

Thank you for your inquiry.

Since the working dilutions of antibodies are not a fixed value but have to optimised by the endusers themselves according to experimental condition, I cannot give you absolute numbers. Even when the pre-diluted antibodies are used neat there might be reasons to use 50ul per slide or maybe even 100ul per slide.

These conditions include: size of the sample, tissue type, lab experience, diluent buffer, species, antigen retrieval, incubation time, etc.

For the products you mentioned I can give a estimate for the pre-diluted antibodies of ca 140- 280slides, for the other antibodies 500 - 100 slides and for the detection kit 150 - 300 slides.

I hope this information is helpful.

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Answer

Thank you for your inquiry.

The protocol for ab64264 reflects the sequence we follow in house and this kit is tested towork useddoing the steps in this order.

Personally I have done the endogenous peroxidase blocking step before antigen retrieval, after antigen retrieval and after the primary antibody and my sense is that it doesn’t matter whThank you for your inquiry.

The protocol for ab64264 reflects the sequence we follow in house and this kit is tested to work used doing the steps in this order.

Personally I have done the endogenous peroxidase blocking step before antigen retrieval, after antigen retrieval and after the primary antibody and my sense is that it doesn’t matter when as long as it is before the HRP step. I do not have any experimental data to support that statement but that is my personal opinion.

Please do not hesitate to contact me with additional questions or concerns.
en as long as it is before the HRP step. I do not have any experimental data to support that statement but that is my personal opinion.

Please do not hesitate to contact me with additional questions or concerns.

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Answer

Thank you for contacting us. Theoratically, this kit can be used for IHC-Fr also. Please do not follow first step in protocol. For other steps please follow the protocol as required. The number of reactions depends on the use of this kit. There is 15 ml solution of each component except DAB chromogen (0.5ml, 50X) which will be suitable for many reactions based on the surface area of tissue sections. Please be advised that 1ml volume informaiton on the datasheet is incoprrect at the moment, we will be updating it shortly. 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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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