Mouse monoclonal [2A9] Anti-Rabbit IgG heavy chain (HRP) (ab99702)

Overview

  • Product name

    Mouse monoclonal [2A9] Anti-Rabbit IgG heavy chain (HRP)
    See all IgG secondary antibodies
  • Host species

    Mouse
  • Target species

    Rabbit
  • Specificity

    ab99702 reacts with the heavy chain of all subclasses of rabbit IgG. Minimal reactivity to human, mouse, rat, goat, bovine, horse, donkey, sheep, guinea pig and chicken serum.

  • Tested applications

    Suitable for: ELISA, WB, IHC-Pmore details
  • Immunogen

    Recombinant fragment corresponding to IgG. Recombinant rabbit IgG Fc fragment

  • Conjugation

    HRP

Properties

Applications

Our Abpromise guarantee covers the use of ab99702 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
ELISA 1/4000 - 1/8000.
WB Use at an assay dependent concentration.
IHC-P 1/8000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Images

  • A rabbit polyconial iNOS to immunoprecipitate HMGB1 and detect the association by immunoblotting of HMGB1 (ab18265). Due to the size of HMGB1, the IgG light chain masked the HMGB1 signal in iNOS immunoprecipitated samples. To avoid the effects from IgG light chain, Mouse monoclonal 2A9 Anti-Rabbit IgG heavy chain (HRP) (ab99702) is chosen to replace the goat anti-rabbit IgG (H&L) HRP-conjugated secondary antibody used.

    As indicated in the image, Mouse monoclonal 2A9 Anti-Rabbit IgG heavy chain (HRP) (ab99702) is shown in the left panel while the goat anti-rabbit IgG (H&L) HRP-conjugated secondary antibody (other source) is shown in the right panel.

  • IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*.

    The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6) for 30 mins. The section was incubated with ab8227, 3 µg/ml overnight at +4°C. An HRP-conjugated secondary (ab99702, 1/8000 dilution) was used for 1hr at room temperature. The section was counterstained with hematoxylin and mounted with DPX.

    The inset negative control image is secondary only at 1/4000 dilution.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Lane 1: Rabbit IgG.

    Lane 2: Rabbit IgG light chains.

    Lane 3: Rabbit IgG heavy chains.

  • ELISA plate was coated with purified rabbit IgG and IgM.

    Immunoglobulins were detected with serially diluted ab99702.

References

This product has been referenced in:

  • Shan Z  et al. Puerarin promotes the proliferation and differentiation of MC3T3-E1 cells via microRNA-106b by targeting receptor activator of nuclear factor-?B ligand. Exp Ther Med 15:55-60 (2018). Read more (PubMed: 29375675) »
  • Zhao SJ  et al. GIT1 contributes to autophagy in osteoclast through disruption of the binding of Beclin1 and Bcl2 under starvation condition. Cell Death Dis 9:1195 (2018). Read more (PubMed: 30546041) »
See all 16 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Western blot
We used rabbit polyconial iNOS to immunoprecipitate HMGB1 and detect the association by immunoblotting of HMGB1 (ab18265). Due to the size of HMGB1, the IgG light chain masked the HMGB1 signal in iNOS immunoprecipitated samples. To avoid the effects from IgG light chain, Mouse monoclonal 2A9 Anti-Rabbit IgG heavy chain (HRP) (ab99702) is chosen to replace the goat anti-rabbit IgG (H&L) HRP-conjugated secondary antibody from Cell Signaling.

As indicated in the image, Mouse monoclonal 2A9 Anti-Rabbit IgG heavy chain (HRP) (ab99702) is shown in the left panel while the goat anti-rabbit IgG (H&L) HRP-conjugated secondary antibody from Cell Signaling is shown in the right panel.

Abcam user community

Verified customer

Submitted Sep 02 2014

Application
Immunoprecipitation
Myc tagged 25kDa protein probed with a Myc polyclonal antibody and a secondary Rabbit heavy chain specific antibody after Immunoprecipitation Lane 6 is the negative control for IP. The specific band appears at the right size.

Abcam user community

Verified customer

Submitted Feb 28 2014

Answer

Thank you for contacting us.

ab73752, ab99697, ab99699 and ab99702 have been tested to detect their specific proteins by Western blot under reducing and denatured conditions. Therefore they can recognise their targets in denatured conditions.

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 contacting us.



The concentration for the current lot (GR68281) of ab99702,Mouse monoclonal [2A9] Secondary Antibody to Rabbit IgG - heavy chain (HRP), is 0.35mg/mL.





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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