Overview

  • Product name

  • Description

    Rabbit polyclonal to GAD67
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Cat, Dog, Pig, Zebrafish
  • Immunogen

    Recombinant fragment, corresponding to a region within amino acids 5 and 176 of GAD67.

  • Positive control

    • H1299, HeLa, HepG2, Molt-4, Raji and NIH-3T3 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab97739 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
WB 1/500 - 1/3000. Predicted molecular weight: 67 kDa.
ICC/IF 1/100 - 1/200.

Target

  • Function

    Catalyzes the production of GABA.
  • Tissue specificity

    Isoform 3 is expressed in pancreatic islets, testis, adrenal cortex, and perhaps other endocrine tissues, but not in brain.
  • Involvement in disease

    Defects in GAD1 are the cause of cerebral palsy spastic quadriplegic type 1 (CPSQ1) [MIM:603513]. A non-progressive disorder of movement and/or posture resulting from defects in the developing central nervous system. Affected individuals manifest symmetrical, non-progressive spasticity and no adverse perinatal history or obvious underlying alternative diagnosis. Developmental delay, mental retardation and sometimes epilepsy can be part of the clinical picture.
  • Sequence similarities

    Belongs to the group II decarboxylase family.
  • Information by UniProt
  • Database links

  • Alternative names

    • 67 kDa glutamic acid decarboxylase antibody
    • CPSQ1 antibody
    • DCE1 antibody
    • DCE1_HUMAN antibody
    • EC 4.1.1.15 antibody
    • FLJ45882 antibody
    • GAD 1 antibody
    • GAD 67 antibody
    • GAD antibody
    • GAD-67 antibody
    • GAD1 antibody
    • Glutamate decarboxylase 1 (brain, 67kDa) antibody
    • Glutamate decarboxylase 1 antibody
    • Glutamate decarboxylase 1 brain 67kD antibody
    • Glutamate decarboxylase 1 brain 67kDa antibody
    • Glutamate decarboxylase 67 kDa isoform antibody
    • Glutamate decarboxylase, brain, 67-KD antibody
    • OTTHUMP00000041054 antibody
    • OTTHUMP00000041055 antibody
    • OTTHUMP00000228572 antibody
    • SCP antibody
    see all

Images

  • Anti-GAD67 antibody (ab97739) at 1/500 dilution + HepG2 whole cell lysate at 30 µg

    Predicted band size: 67 kDa



    7.5% SDS PAGE
  • Anti-GAD67 antibody (ab97739) at 1/1000 dilution + NIH-3T3 whole cell lysate at 30 µg

    Predicted band size: 67 kDa



    7.5% SDS PAGE
  • Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using GAD67 antibody (ab97739) at 1/200 dilution. Lower image was also merged with DNA probe.

References

This product has been referenced in:

  • Nobili A  et al. Ambra1 Shapes Hippocampal Inhibition/Excitation Balance: Role in Neurodevelopmental Disorders. Mol Neurobiol 55:7921-7940 (2018). Read more (PubMed: 29488136) »
  • Ding Y  et al. Changes in GABAergic markers accompany degradation of neuronal function in the primary visual cortex of senescent rats. Sci Rep 7:14897 (2017). WB . Read more (PubMed: 29097694) »
See all 6 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Squirrel Tissue sections (Brain, preoptic area)
Antigen retrieval step
None
Permeabilization
No
Specification
Brain, preoptic area
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Nov 20 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Brain)
Permeabilization
Yes - Triton-X
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 21°C
Fixative
Paraformaldehyde

S Jarrin

Verified customer

Submitted Jul 11 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain Tissue 40um slices)
Antigen retrieval step
None
Permeabilization
Yes - Triton X in PBS
Specification
Brain Tissue 40um slices
Blocking step
Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 24 2015

Question

Here are the answers to your questions:

What kind of samples do you have (species, cells or tissues, etc)?

We are performing standard immunofluorescence IHC on mouse and rat brain tissue. Animals were perfused with 4% paraformaldehyde and tissue was sectioned in a cryostat at 35um thickness.

Could you describe the results you've seen with the antibody and the protocol that was used?

A standard IHC technique was used:

Tissue was rinsed in TBS buffer, incubated with sodium borohydride for 30minutes, rinsed, incubated in blocking solution (gelatin, NGS), incubated in primary antibody for 2 nights, rinsed, incubated in Alexa 555 donkey anti rabbit secondary antibody, and rinsed again before mounting. No triton X was used.

Upon initial testing the antibody gave what appeared to be strong staining of multiple cell bodies in the brainstem. The cell population stained, however, seemed somewhat large to correspond to the previously identified sparse GABAergic population of that region.

Which amino acids does the control peptide correspond to and what is the protocol for blocking the antibody?

The control peptide used was the entire GAD67 protein, purchased from alpha-diagnostics. Control peptide was used at a concentration of 25x that of the primary antibody and the two were incubated overnight at 4*C prior to addition to the tissue.

After being informed that the primary antibody was not to the originally described sequence, we ran two specificity experiments: 1. Block by 25x GAD67 control peptide- no block was achieved; 2. Staining was tested in the hippocampus, where there are well characterized and anatomically defined GABAergic neuronal populations. The staining pattern we found with this antibody did not correspond with this anatomy in that many non-GABAergic cell populations also stained positive. (In the hippocampus only interneurons are GABAergic, the pyramidal cell population is not.) Both of these control experiments suggest that this antibody is staining non-GABAergic populations.

We would prefer to receive a credit refund for the three vials. We can certainly return the two unopened vials.
Thanks very much,

Read More
Answer

Thank you very much for your reply and for sending the additional information.

We have only tested the antibody in Western blot and in IF staining on fixed cells, so we aren't sure at this time whether it is suitable for use on perfused-fixed tissue. However I understand that the specificity of the antibody is in question, especiallydue tothe immunogen update. I've issued the credit notes below, and the full purchase prices will be refunded back to your credit card pending approval of our accounting department:

***

If you have any questions about these refunds, please contact our accounting department at mailto:us.credits@abcam.com and reference the credit note numbers. You do not need to return the unopened vials.

I am sorry for the inconvenience regarding ab97739, and if there is anything else that we can do for you please let me know. I'll be happy to help.

Read More

Answer

Thank you very much for contacting us and letting us know about the trouble with this antibody.

I am sorry to hear that the results have not been satisfactory when using ab97739. We didupdate the immunogen region that was previously listed on our datasheet, however this antibody is raised against GAD67 andwe expect it tobe specific for this protein.

Could you describe the results you've seen with the antibody and the protocol that was used? What kind of samples do you have (species, cells or tissues, etc)? Which amino acids does the control peptide correspond to and what is the protocol for blocking the antibody?

We do fully guarantee this antibody to work as specificed on the datasheet in Western blot and immunocytochemistry on human and mouse samples, and if it is not working correctly we can send a replacement or alternatively issue a credit or refund if you prefer. If you would be interested in one of these options, could you please send me your order number or PO number?

I look forward to hearing from you. Please let me know if you have further questions or if there is anything else that we can do for you.

Read More

Question
Answer

DISCOUNT CODE: 100%ABR-T76WH
Expiration date: 08-11-2012

I am very pleased to hear you would like to use ab97739 inIHC-FoFr and send us an image of your results.

This code will give you 1 free [PRIMARY ANTIBODY] before the expiration date. To redeem this offer, please submit an Abreview with the image forIHC-FoFr 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: www.abcam.com/Abreviews.

The code will be active once the Abreview with the image 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. Remember if the product does not work you will be covered by our Abpromise guarantee and eligible for a full refund or replacement.

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.

PS: The ab97739 is a rabbit polyclonal ab so you willneed anti rabbit secondary antibody.

Read More

Question
Answer

Thank you for your inquiry.

Unfortunately, the blocking peptide forab97739 is not available for purchase.

Please let me know if there is anything else I can help you with.

Read More

Answer

Thank you for contacting us.

Your credit note ID for the refund is xxx.

I am sorry that this antibody did not have the correct immunogen informationon the datasheet, which we have corrected in the meantime. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

Read More

Question
Answer

Thank you for contacting us.

I have had the opportunity to inquire about the availability of the GAD76 blocking peptide as you had requested. Unfortunately I have been told that this is not presently available. While this control is not available we do carry a number of positive control lysates recommended on the datasheet such as HeLa, HepG2, Molt-4, Raji and NIH-3T3 cells. I have added links to these controls below:

HeLa: https://www.abcam.com/HeLa-Human-epithelial-carcinoma-cell-line-Nuclear-Lysate-ab14655.html
HepG2: https://www.abcam.com/HepG2-Human-hepatocellular-liver-carcinoma-cell-line-Whole-Cell-Lysate-ab7900.html
Molt-4: https://www.abcam.com/MOLT4-Human-acute-lymphoblastic-leukemia-cell-line-Whole-Cell-Lysate-ab7912.html
Raji: https://www.abcam.com/Raji-Human-Burkitt-s-lymphoma-cell-line-Whole-Cell-Lysate-tumor-cell-line-ab30124.html
NIH-3T3: https://www.abcam.com/NIH-3T3-Mouse-embryonic-fibroblast-cell-line-Whole-Cell-Lysate-ab7179.html


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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