Overview

  • Product name
    Anti-Germ cell-specific antigen antibody [TRA98]
  • Description
    Rat monoclonal [TRA98] to Germ cell-specific antigen
  • Host species
    Rat
  • Specificity
    In PubMed: 27718356 the epitope of ab82527 has been mapped to a murine-specific 8-amino-acid tandem GE(P/M/S)E(S/T)EAK repeat that occurs 25 times in mouse GCNA1 (aa 167- 383).
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Tissue, cells or virus corresponding to Mouse Germ cell-specific antigen. Cell lysates of adult mouse testes.

  • Positive control
    • Testicular germ cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab82527 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 dilution.
WB 1/5000. Predicted molecular weight: 146 kDa.

Target

  • Relevance
    Detected during development of germ cells and also after birth, but the signal in female disappears by 5 days after birth. The antigen is localized only in the nuclear fraction of testicular germ cells.
  • Alternative names
    • GENA antibody
    • Testicular germ cell-specific antigen antibody

Images

  • ab82527 staining Germ cell-specific antigen in mouse postnatal day 7 testes tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 4% paraformaldehyde and blocked with 5% BSA for 45 minutes at 25°C; antigen retrieval was by heat mediation in 0.1M citric acid/sodium citrate buffer. Samples were incubated with primary antibody (1/500 in 1% BSA) for 1 hour at 25°C. A biotin-conjugated goat anti-rat IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • ab82527 staining Germ cell-specific antigen in adult mouse testis tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton and blocked with 3% BSA in PBS for 30 minutes at 20°C. Samples were incubated with primary antibody (1/1000) for 1 hour at 20°C. Ab150165 was used as the secondary antibody, undiluted.

    See Abreview

  • All lanes : Anti-Germ cell-specific antigen antibody [TRA98] (ab82527) at 0.1 µg/ml

    Lane 1 : mouse brain lysate
    Lane 2 : mouse heart lysate
    Lane 3 : mouse intestine lysate
    Lane 4 : mouse kidney lysate
    Lane 5 : mouse liver lysate
    Lane 6 : mouse lung lysate
    Lane 7 : mouse spleen lysate
    Lane 8 : mouse ovary lysate
    Lane 9 : mouse testis lysate

    Lysates/proteins at 100 µg per lane.

    Predicted band size: 146 kDa



    Western blot analysis of various mouse tissues, using ab82527.
    In adult tissues, bands of 60-100 kDa proteins are detected only in the testis. The signals are observed in male and female embryos after embryonic day 12.5. The signal in male is detected during development of germ cells and also after birth, but the signal in female disappears by 5 days after birth. The antigen is localized only in the nuclear fraction of testicular germ cells.
  • Immunohistochemical analysis of mouse fetal ovary tissue, staining Germ cell-specific antigen with ab82527.

    Tissue was fixed with Bouin's fixative, permeabilized with 0.05% Tween 20 and 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200 in 1% BSA and 0.5% Triton X-100 in 1X PBS) for 1 hour at 25°C. An AlexaFluor®488-conjugated goat anti-rat polyclonal IgG (1/200) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Simanjuntak Y  et al. Ebselen alleviates testicular pathology in mice with Zika virus infection and prevents its sexual transmission. PLoS Pathog 14:e1006854 (2018). Read more (PubMed: 29447264) »
  • Niedenberger BA & Geyer CB Advanced immunostaining approaches to study early male germ cell development. Stem Cell Res 27:162-168 (2018). IHC-P ; Mouse . Read more (PubMed: 29475796) »
See all 24 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Testis, adult)
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Specification
Testis, adult
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Fixative
Paraformaldehyde

Mr. Bryan Niedenberger

Verified customer

Submitted Apr 04 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (fibroblast)
Permeabilization
No
Specification
fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
ice-cold acetone

Abcam user community

Verified customer

Submitted Jul 07 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate
Sample
Mouse Tissue sections (Postnatal day 7 mouse testes)
Specification
Postnatal day 7 mouse testes
Permeabilization
No
Fixative
4% PFA

Abcam user community

Verified customer

Submitted Dec 29 2014

Question
Answer

Thank you for your inquiry and your patience with this reply.
For ab82527, it should be IgG2a, although we do not do any validation to verify that. The clone TRA98 recognizes a mouse testicular germ cell-specific antigen (1). In adult tissues, bands of 60-100 kDa proteins are detected only in the testis by Western blotting analysis with TRA98 (Fig on datasheet). The signals are observed in male and female embryos after embryonic day 12.5. The signal in male is detected during development of germ cells and also after birth, but the signal in female disappears by 5 days after birth. The antigen is localized only in the nuclear fraction of testicular germ cells and this antibody can be used for immunohistochemical staining of testicular germ cells (1, 2). The hybridoma producing TRA98 was obtained by immunization of a rat with the cell lysate of adult mouse testis by the group of Prof. Yoshitake Nishimune of Osaka University (1). This product was purified from serum-free culture medium of the hybridoma clone TRA98 by proprietary chromatography processes under mild conditions.
References: This product has been used in the following publications.
1.Tanaka H et al ” A germ cell-specific nuclear antigen recognized by a monoclonal antibody raised against mouse testicular germ cells” Int J Androl 20: 361-366 (1997) PMID: 9568529
2. Ohta H et al “Regulation of proliferation and differentiation in spermatogonial stem cells: the role of c-kit and its ligand SCF” Development 127: 2125-2131 (2000) PMID: 10769236
I think the other antibody you were looking at may have been clone TRA54 (IgM), ab92286. It is a different clone from TRA54 in that they recognize different proteins, although both target proteins are specific for haploid sperm cell-specific proteins. Dr. Tanaka who provided these clones, says the genes that code for the target proteins will be shortly published.
https://www.abcam.com/haploid-sperm-cell-specific-antigen-antibody-tra54-ab92286.html
I hope this information helps. Please contact us with any other questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (fetal ovary)
Specification
fetal ovary
Fixative
Bouin's fixative
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer
Permeabilization
Yes - 0.05% Tween 20 and 0.1% Triton X-100 in blocking serum
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 06 2012

Question
Answer

DISCOUNT CODE: ***
Expiration date: February 15th, 2012
Value: ***

I am very pleased to hear you would like to accept our offer and test ab82527 in IHC-P. This code will give you *** off your next 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. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: https://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: https://www.abcam.com/abtrial.

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Answer

Thank you very much for your email.
I am happy to arrange a credit note or refund. However I would need more information about the problem customer experienced. Our customers feedback and own tested data show that the antibody detects Germ cells very nice so I would be interested in to know in which type of experiment the antibody wasn't working.
Many thanks for your cooperation in this case. I will look forward to hearing from you soon.

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Answer

Thank you very much for your valuable feedback.

We would be very grateful if you could submit an Abreview about this product via the online product datasheet. We always encourage customers to send their results back to us.

To find out more about our Abreview system, please see the following webpage:

https://www.abcam.com/abreviews

We hope to receive your Abreview shortly. Thank you for your time and effort.

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Answer

Thank you for contacting us. To answer your questions: 1) Yes, JLP is the same as SPAG9. For more alternative names, please check the protein database entry for the mouse protein: http://www.uniprot.org/uniprot/Q58A65 2)References are listed in the reference tab at the top of the datasheet. I added them here as well: i) Bagheri-Fam S et al. Defective survival of proliferating Sertoli cells and androgen receptor function in a mouse model of the ATR-X syndrome. Hum Mol Genet 20:2213-24 (2011). PubMed: 21427128 ii) Ohta H et al. Regulation of proliferation and differentiation in spermatogonial stem cells: the role of c-kit and its ligand SCF. Development 127:2125-31 (2000). IHC-Fr; Mouse. PubMed: 10769236 iii) Tanaka H et al. A germ cell-specific nuclear antigen recognized by a monoclonal antibody raised against mouse testicular germ cells. Int J Androl 20:361-6 (1997). WB; Mouse. PubMed: 9568529 3) The WB image was obtained fromthe supplying lab. I hope this information is helpful to you. If you are in need of any other product that you might require for your experiments (e.g. secondary antibodies, controls), please let me know as well. Please do not hesitate to contact us if you need any more advice or information.

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