Anti-GABA A Receptor alpha 2/GABRA2 antibody (ab72445)

Reviews (1)Q&A (3)References (13)
Western blot - Anti-GABA A Receptor alpha 2/GABRA2 antibody (ab72445)

    Key features and details

    • Rabbit polyclonal to GABA A Receptor alpha 2/GABRA2
    • Suitable for: WB
    • Reacts with: Mouse
    • Isotype: IgG

    Overview

    • Product name

      Anti-GABA A Receptor alpha 2/GABRA2 antibody
      See all GABA A Receptor alpha 2/GABRA2 primary antibodies

    • Description

      Rabbit polyclonal to GABA A Receptor alpha 2/GABRA2

    • Host species

      Rabbit

    • Specificity

      ab72445 is specific for GABA A Receptor alpha 2/GABRA2

    • Tested Applications & Species

      Application Species
      WB
      Mouse
      See all applications and species data

    • Immunogen

      Fusion protein corresponding to Rat GABA A Receptor alpha 2/GABRA2 (intracellular).

    • Positive control

      • rat brain lysates

    • General notes

       This product was previously labelled as GABA A Receptor alpha 2

       

    Properties

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab72445 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Guaranteed

    Tested applications are guaranteed to work and covered by our Abpromise guarantee.

    Predicted

    Predicted to work for this combination of applications and species but not guaranteed.

    Incompatible

    Does not work for this combination of applications and species.

    Application Species
    WB
    Mouse
    All applications
    Cow
    Dog
    Human
    Non human primates
    Zebrafish
    Application Abreviews Notes
    WB
    1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
    Notes
    WB
    1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).

    Target

    • Function

      GABA, the major inhibitory neurotransmitter in the vertebrate brain, mediates neuronal inhibition by binding to the GABA/benzodiazepine receptor and opening an integral chloride channel.

    • Sequence similarities

      Belongs to the ligand-gated ion channel (TC 1.A.9) family. Gamma-aminobutyric acid receptor (TC 1.A.9.5) subfamily. GABRA2 sub-subfamily.

    • Cellular localization

      Cell junction > synapse > postsynaptic cell membrane. Cell membrane.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • GABA A receptor subunit alpha 2 antibody
      • GABA antibody
      • GABA(A) receptor subunit alpha 2 antibody
      • GABA(A) receptor subunit alpha-2 antibody
      • GABR A2 antibody
      • GABRA 2 antibody
      • GABRA2 antibody
      • GABRA2 protein antibody
      • Gamma aminobutyric acid (GABA) A receptor alpha 2 antibody
      • Gamma aminobutyric acid A receptor alpha 2 antibody
      • Gamma aminobutyric acid receptor alpha 2 subunit antibody
      • Gamma aminobutyric acid receptor subunit alpha 2 antibody
      • Gamma-aminobutyric acid receptor subunit alpha-2 antibody
      • GBRA2_HUMAN antibody
      see all

    Images

    • Western blot - Anti-GABA A Receptor alpha 2/GABRA2 antibody (ab72445)
      Western blot - Anti-GABA A Receptor alpha 2/GABRA2 antibody (ab72445)
      All lanes : Anti-GABA A Receptor alpha 2/GABRA2 antibody (ab72445) at 1/1000 dilution

      Lane 1 : Wild type Mouse brain lysates
      Lane 2 : GABA A Receptor alpha 2/GABRA2 knockout mouse brain lysate

      Predicted band size: 51 kDa
      Observed band size: 51 kDa

    References (13)

    Publishing research using ab72445? Please let us know so that we can cite the reference in this datasheet.

    ab72445 has been referenced in 13 publications.

    • Hosseini M  et al. Simultaneous intrathecal injection of muscimol and endomorphin-1 alleviates neuropathic pain in rat model of spinal cord injury. Brain Behav 10:e01576 (2020). PubMed: 32189472
    • Zhang Y  et al. MeCP2 in cholinergic interneurons of nucleus accumbens regulates fear learning. Elife 9:N/A (2020). PubMed: 32420873
    • Wang XS  et al. Anxiolytic effects of Formononetin in an inflammatory pain mouse model. Mol Brain 12:36 (2019). PubMed: 30961625
    • Zhang K  et al. CB1 agonism prolongs therapeutic window for hormone replacement in ovariectomized mice. J Clin Invest 129:2333-2350 (2019). PubMed: 31063987
    • Pawar HN  et al. Does acute heat stress differentially-modulate expression of ionotropic neurotransmitter receptors in the RVLM of young and aged F344 rats? Neurosci Lett 687:223-233 (2018). PubMed: 30287307
    View all Publications for this product

    Customer reviews and Q&As

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    1-4 of 4 Abreviews or Q&A

    Immunohistochemistry (Frozen sections) abreview for Anti-GABA A Receptor alpha 2 antibody

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain, Frontal cortex)
    Permeabilization
    Yes - Triton X-100
    Specification
    Brain, Frontal cortex
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 0.2% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 29 2017

    Question

    Can you please provide the exact immunogen sequence? Is it extracellular?

    Read More
    Answer

    Thank you for contacting Abcam regarding the immunogen information for ab72445 and ab103707.

    Regarding ab103707, the immunogen was generated from a peptide within aa53-83, corresponding to an extracellular domain of the protein. The specific aa sequence is proprietary and unfortunately I do not have any additional information.

    The immunogen used to generate ab72445 is a peptide that corresponds to a cytoplasmic region of the protein.

    I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

    Read More

    Question


    Thanks for getting back to me so quickly.
    Here are the answers to the questions you asked:
    1 - Are you boiling your samples? If so for how long?
    I did not "boil" my samples but heated them up at 85C for 10 mins.
    2 - What percentage of SDS gels are you running (8, 12.5, 15%)?
    The images I sent are from a 12.5% gel. I also tried 8%, with almost identical results.
    3 - At what voltage do you run your SDS gel?
    120V for a little over 2 hours
    4 - What is the recipe of your transfer buffer and how long do you transfer for?
    Transfer buffer (10X):
    30.3g Tris base
    144.1g glycine
    Top up to 1000mL with ddH2O
    To make 1x:
    100mL 10x stock
    500mL ddH2O
    200mL methanol
    Top up to 1000mL with ddH2O
    I keep the 10x stock at 4C and add cold ddH2O to make sure that the buffer is cold.
    I transferred at 100V for 1.5hrs
    5 - How much total protein are you loading into each well?
    30ug
    Thanks again!

    Read More
    Answer

    Thank you for your reply.

    There are few protocol variations that may help improve the results that your are getting:

    1 - Do not heat/boil your samples. When looking at transmembrane proteins, it is advisable to not heat the sample as the heating will cause the proteins to 'clump' together, thereby affecting its ability to migrate through the gel.

    2 - When running your gel, I would run the gel for 30 minutes or so at 90V (or until the samples are through the stacking layer) and then increase the voltage to 120v until it is finished. That should help improve the bands that are being resolve on the membrane.

    3 - You could try doing an overnight transfer of the gel. You would do it at 25V for at least 14hrs and I would do it in the fridge, so that your buffer stays cold (this is not necessary, but could help).

    For the size of bands that you are probing for, a 12.5% gel is what I would recommend and the transfer buffer you are using is what I would use.

    Both antibodies (ab103707 & ab72445) are covered under our Abpromise for six months and are guaranteed to work in western blot. . If we cannot resolve the issue you are having with these antibodies then I would be happy to either send a replacement antibody or to process a refund.

    Please let me know if there is anything else I can help you with, or if the protocol variations do not prove to be successful.

    Read More

    Question

    first few trials have unfortunately given weird outcomes, and because of lack of experience, I have no clue what the reason might be. I am attaching pictures from one of the trials; the top blot is alpha5 and the lower one is alpha 2 in both pictures. The beta actin blot is from the same blots, which I stripped using the mild stripping protocol from your website the next day and reprobed for actin. The samples were whole brain samples from mice which I lysed in 2% SDS in 60mM Tris (pH 6.8). I did another round where I added leupeptin, pepstatin, antipain and EDTA to the lysis buffer and it gave pretty much identical results. I blocked at RT for one hour in 5% milk in TBST before incubation in primary antibody and dissolved the primary antibodies in this solution too. I incubated in primary overnight at 4C. The next morning I did 3 x 20min washes in TBST, followed by one hour incubation in secondary antibody dissolved in 5%milk in TBST, and another 3 x 20min washes in TBST. The antibodies and concentrations I used are as follows:
    -Alpha 5:
    Primary: ab103707, 1:300
    Secondary: ab97200, 1:5000
    -Alpha2:
    Primary: ab72445, 1:1000
    Secondary: ab97200, 1:5000
    -bActin:

    As you can see from the blots, the problems are two-fold. First of all, the quality in general is terrible, things seem to be kind of leaking out of the lanes and not giving me real bands but these smudges. I have no idea what may have caused it and haven’t seen anything fitting this description in your WB troubleshooting documents online either. The second problem is the results I am getting with the alpha 2 and alpha 5 antibodies. While the quality is still bad, I seem to be at least getting a band at the correct size for the actin, whereas the bands for alpha2 and alpha 5 don’t make any sense. For alpha 2, I don’t even have anything at the expected size and I get this really large band on top every time I run it. For alpha 5, it looks like I might at least have something at the right size in addition to lots of other stuff, but this is not true, because the last 2 lanes are from alpha 5 global knockout animals and they still have this band. I would really appreciate any help you can provide. If you could make suggestions regarding how to improve the general quality first, I can certainly run more samples and send more pictures. Also, please let me know if you need more information regarding my protocol.

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    Answer

    Thank you for contacting Abcam.

    I would be more than happy to help out in any way that I can. I just have a few more protocol questions for you:

    1 - Are you boiling your samples? If so for how long?

    2 - What percentage of SDS gels are you running (8, 12.5, 15%)?

    3 - At what voltage do you run your SDS gel?

    4 - What is the recipe of your transfer buffer and how long do you transfer for?

    5 - How much total protein are you loading into each well?

    I look forward to your reply and helping you resolve these issue that you are having.

    Read More

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