Overview

  • Product name
    BrdU Immunohistochemistry Kit
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Product overview

    ab125306 is a histochemical staining kit for the detection and localization of bromodeoxyuridine incorporated into newly synthesized DNA of actively proliferating cells.


    The material in ab125306 is sufficient to run 50 slides. The average test area is defined as a circle around the tissue with an approximate diameter of 2 centimeters.


    Control Slides: Mouse intestinal tissue pulsed with BrdU ab129956 


     

  • Notes

    *Trypsin is only required if using formalin fixed tissues. If the tissues are fixed in alcohol, trypsin digestion is not required. Please note that a quenching solution (hydrogen peroxide (30% solution)) is not provided in this kit.

     

     

  • Tested applications
    Suitable for: IHC-Fr, IHC-Pmore details

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab125306 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-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Images

  • Formalin-fixed, paraffin embedded mouse intestinal tissue sections stained using ab125306.
  • Formalin-fixed, paraffin embedded mouse intestinal tissue sections stained using ab125306.
  • Formalin-fixed, paraffin embedded mouse intestinal tissue sections stained using ab125306.

Protocols

References

This product has been referenced in:
  • Norum JH  et al. The tankyrase inhibitor G007-LK inhibits small intestine LGR5+stem cell proliferation without altering tissue morphology. Biol Res 51:3 (2018). Mouse . Read more (PubMed: 29316982) »
  • Gou Y  et al. Protein Arginine Methyltransferase PRMT1 Is Essential for Palatogenesis. J Dent Res N/A:22034518785164 (2018). Read more (PubMed: 29986157) »
See all 9 Publications for this product

Customer reviews and Q&As

1-4 of 4 Q&A

Answer

The detection antibody in ab125306 is an affinity purified sheep polyclonal conjugated to biotin. This antibody is not available for purchase separately outside of the kit.

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Answer

Following the staining protocol on page 11 of the protocol for ab125306, with frozen tissue section you would skip step 1 "Deparaffinization" and start with step 2.1. You will also skip step 2.2, as you do not need to perform antigen retrieval unless your tissues were formalin fixed. If you have PFA perfusion fixed, frozen tissue you will most likely still need to included step 2.2, but if you used a fixative such as methanol, ethanol, or acetone you can skip step 2.2.

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Question
Answer

To answer your questions about ab125306:
1) For suspension cells – is cytospin the only preparation method for suspension cells?
It is possible to prepare slides by preparing a 'slurry' suspension of cells and air drying them onto slides. Followed by dehydration fixing with 70 % etoh and processing. Cell morphology however is compromised for non adherent cells.
2) Other antigen retrieval methods may be used, should these be PIER as well or would HIER techniques be okay?
Proteases are required to reveal epitopes for the IHC BrdU kit.
3) Why can’t tissues/cells be rinsed after blocking step? If a customer accidentally does this, can they repeat just the blocking step and proceed with the protocol or do they need to repeat the denaturing step as well.
Rinsing after the blocking step may slightly increase background but shouldn't be 'terminal'. It is not required to repeat the denaturing step.
4) What is the purpose of the ethanol incubation step followed by xylene (this is after the counterstain) (mentioned on page 4)?
Ethanol {100 - 70% followed by PBS) is used after xylene to rehydrate the tissue. The procedure for dehydrating samples (prior to cover slipping) is the reverse. Incubate slides in 90% ethanol for 30 seconds, 100% ethanol for 30 seconds and xylene for 30 seconds (2 times each). Add 1-2 drops of mount- ing media and coverslip.

5) For paraffin sections, what are the guidelines for BrdU treatment of an animal [a) how much BrdU per animal; b) how long of a treatment with BrdU]?
Protocol attached.
6) For BrdU staining of cultured cells and cell suspensions: Under section A 3. Suitable slides are mentioned. Are any specific slides recommended? Do they need to be coated?
'Appropriate' slides refers to coated slides. Coated slides are required.
7) Should I want to perform an isotype control; what is the species, isotype and concentration of the detector ab?
It wil be difficult to use an appropriate isotype control - the concentrations of antibody and detector reagents in the kit are LOT specific and vary. A no BrdU control will provide a suitable control as all other reagents are used (except BrdU).
8) Will this product work with yeast?
The assay should work in any cell type that incorporates BrdU.
9) Can TBS be used in lieu of PBS?
TBS can be used however the assay has been optimized for use with PBS and we have no in house experience using TBS in the assay.

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Question
Answer

The catalogue number we have is ab129956.

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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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