Overview

  • Product name
    Anti-GABA A Receptor alpha 1 antibody
    See all GABA A Receptor alpha 1 primary antibodies
  • Description
    Rabbit polyclonal to GABA A Receptor alpha 1
  • Host species
    Rabbit
  • Specificity
    This antibody is specific for the GABA A Receptor a1 subunit.
  • Tested applications
    Suitable for: IHC-P, WB, IP, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
    Predicted to work with: Cow, Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    QPSQDELKDNTTVFT-C

    , corresponding to N terminal amino acids 1-15 of the mature Rat GABA A Receptor alpha 1 containing a cysteine at the C terminus for linkage to a carrier protein.

  • Positive control
    • WB and IP: Rat brain microsomal tissue extract

Properties

Applications

Our Abpromise guarantee covers the use of ab33299 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 1/500. PubMed: 20650017
WB 1/10000. Predicted molecular weight: 52 kDa.
IP Use at an assay dependent concentration. 5ul immunoprecipitates the GABA A Receptor a1 subunit in rat brain microsomal preparation RIPA lysate.
IHC-Fr Use at an assay dependent concentration. Note: use 4% para glutaraldehyde fixed frozen sections permeabilized with Triton X100.

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.
  • Involvement in disease
    Defects in GABRA1 are the cause of childhood absence epilepsy type 4 (ECA4) [MIM:611136]. A subtype of idiopathic generalized epilepsy characterized by onset at age 6-7 years, frequent absence seizures (several per day) and bilateral, synchronous, symmetric 3-Hz spike waves on EEG. During adolescence, tonic-clonic and myoclonic seizures may develop. Absence seizures may either remit or persist into adulthood.
    Defects in GABRA1 are the cause of juvenile myoclonic epilepsy type 5 (EJM5) [MIM:611136]. A subtype of idiopathic generalized epilepsy. Patients have afebrile seizures only, with onset in adolescence (rather than in childhood) and myoclonic jerks which usually occur after awakening and are triggered by sleep deprivation and fatigue.
  • Sequence similarities
    Belongs to the ligand-gated ion channel (TC 1.A.9) family. Gamma-aminobutyric acid receptor (TC 1.A.9.5) subfamily. GABRA1 sub-subfamily.
  • Cellular localization
    Cell junction > synapse > postsynaptic cell membrane. Cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ECA4 antibody
    • EIEE19 antibody
    • EJM antibody
    • EJM5 antibody
    • Gaba receptor alpha 1 polypeptide antibody
    • GABA(A) receptor antibody
    • GABA(A) receptor subunit alpha 1 antibody
    • GABA(A) receptor subunit alpha-1 antibody
    • GABA(A) receptor, alpha 1 antibody
    • GABRA 1 antibody
    • GABRA1 antibody
    • Gamma aminobutyric acid (GABA) A receptor alpha 1 antibody
    • Gamma aminobutyric acid A receptor alpha 1 antibody
    • Gamma aminobutyric acid receptor subunit alpha 1 antibody
    • Gamma aminobutyric acid type A receptor alpha1 subunit antibody
    • Gamma-aminobutyric acid receptor subunit alpha-1 antibody
    • GBRA1_HUMAN antibody
    see all

Images

  • Anti-GABA A Receptor alpha 1 antibody (ab33299) at 1/10000 dilution + rat brain microsomal preparation tissue lysate at 10 µg

    Predicted band size: 52 kDa
    Observed band size: 51 kDa
    why is the actual band size different from the predicted?



    Proteins were visualized using an IgG (HRP) donkey anti-rabbit and a chemiluminescence detection system.

  • Immunohistochemistry analysis of formalin fixed paraffin embedded rat hippocampus tissue prepared using heat-induced epitope retrieval (HIER) labeling GABA A Receptor alpha 1 using ab33299 at 1/500 dilution. Secondary antibody was an anti-rabbit antibody and HRP-DAB.

  • Immunoprecipitation analysis: 1 mg of RIPA lysate of rat brain microsomes
    was immunoprecipitated with 5 uL of ab33299. Arrow indicates GABAA
    Receptor a1 (~50kD).

  • ab33299 staining GABA A Receptor alpha 1 in human retinal tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed with 4% paraformaldehyde, blocked with 10% donkey serum for 20 minutes at 24°C, then incubated with ab33299 at a 1/100 dilution for 16 hours at 4°C. The secondary antibody used was an Alexa-Fluor 488 (green) conjugated donkey anti-rabbit polyclonal, used at a 1/1000 dilution. Counterstained with Hoechst 33258 (blue).

    IPL - inner plexiform layer
    INL - inner nuclear layer
    OPL - outer plexiform layer
    ONL - outer nuclear layer
    INS - inner nuclear segments

    See Abreview

References

This product has been referenced in:
  • Tabouy L  et al. Dysbiosis of microbiome and probiotic treatment in a genetic model of autism spectrum disorders. Brain Behav Immun N/A:N/A (2018). Read more (PubMed: 29787855) »
  • Huang X  et al. Overexpressing wild-type ?2 subunits rescued the seizure phenotype in Gabrg2+/Q390X Dravet syndrome mice. Epilepsia 58:1451-1461 (2017). Read more (PubMed: 28586508) »
See all 14 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Cell (SH-SY5Y))
Permeabilization
Yes - Triton X-100 (0'25%)
Specification
Human Cell (SH-SY5Y)
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 30 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cortical)
Loading amount
20 µg
Specification
Cortical
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Dr. Martin Broadstock

Verified customer

Submitted Aug 16 2012

Question
Answer

Merci de nous avoir contactés.
Voici les liens vers les guides et posters qui je l'espère vous seront utiles:
> DyLight® conjugated secondary antibodies : https://docs.abcam.com/pdf/general/DyLight_flyer.pdf
> A Guide to Fluorochromes : https://docs.abcam.com/pdf/immunology/fluorochrome_guide.pdf
> How to choose the right secondary antibody : https://www.abcam.com/index.html?pageconfig=resource&rid=12825
> How to choose a conjugate : https://www.abcam.com/index.html?pageconfig=resource&rid=12922
J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

Read More

Question

Hi Tech Support, how are you?

Would you be able to help us with the following enquiry please:

Customer’s question:

I have some questions for you regarding the above antibody.

I am working with guinea pig protein brain samples. I used Invitrogen PARIS kit that allows RNA and protein isolation from the same sample simultaneously. In this kit, after adding the lysis buffer and homogenising the brain sample, the homogenised sample can be immediately be kept aside without any further action for protein analysis. I am using B-actin as reference for my western blot analysis. I am trying to measure a lot of samples (control vs treated vs time-points) and I am then using ImageJ to analyse the densitometry of my bands relative to B-actin to get a relative protein expression value.

For my western blot protocol this is what I do:

Measure protein conc. using BCA kit (high conc.)

Use 15ug protein per sample (dilute with water and 6x loading dye added). Load a total of 12uL.

Heat at 95 degrees for 5 mins

Run on Biorad TGX pre-made gel at 300V for 20mins

Transfer using Biorad TGX transfer system for 5mins

Ponceau S for 5 mins (all bands good transfer)

Block 1 hr in 5% skim milk in TBST

Primary ab overnight in blocking buffer (1:800 for gaba a antibody) at 4 degrees in dark cold room

Next day, wash 3x for 5 mins in TBST

Sec ab (anti rabbit, 1:10,000) for 1 hr at room temp in blocking buffer

Wash 3x for 5 mins in TBST

Use ECL for detection (from Milipore)

Detect bands using Biorad ECL transilluminator

Previously, I have combined the antibodies of GABA A and B-actin in the same solution for primary antibody incubation and use anti-mouse and anti-rabbit in the same secondary antibody incubation on the blot (detecting two proteins at the same time on the same blot). From these results, I got two very distinct bands, one at the ˜48kDa area (which I thought is the B-actin band) and a very distinct one at ˜75kDa (which I assumed is the GABA-A band).

Of course, looking at the data sheet, this did not make sense as GABA-A is 52-55kDa. So I did not understand why I was seeing a very distinct band at the 75kDa area. I repeated this many many times and for many many samples and kept getting the same results, though each time when I quantified the intensity of the assumed GABA-A bands, it changed each time giving me highly variable results.

So then, I separated out the antibodies and did them individually. After Ponceau S staining, I cut the membrane up one 50kDa and below, and the other is the top half (containing the 75kDa mark). I made different primary ab solutions and incubated them separately. B-actin came out perfectly but there was nothing for the top half of the membrane (with GABA-A antibody).

I then repeated the whole thing again from the start, using the same protein samples except this time, I did purely GABA-A antibody alone without cutting the membrane up at all. Nothing came up, not even at the 52-55kDa area.

Now of course I am stuck. I reordered a new antibody (ab33299) but I am now considering trying from a different company or getting a positive sample in like HeLa or HepG2 cell line to test if the GABA-A antibody works.

Please Let me know what I can try next or if I should try a different GABA A antibody or a positive control?

Please advise!

Read More
Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

You are correct in thinking the band expected for the GABA A Receptor alpha 1 would be around 52 kDa. Unfortunately, we have not tested this antibody with guinea pig samples as yet and cannot therefore say conclusively that it will be able to cross react. I would therefore say that using a positive control would be very worthwhile. However, I don't think the HeLa or HepG2 would be suitable as this is a protein mainly expressed in the brain.

Do you have any access to human, rat or mouse brain lysates? If you do, I would suggest trying these.

We do not currently have any anti-GABA A Receptor alpha 1 antibodies which have been shown to detect the guinea pig protein but I can checkwhich antibodies may be the most appropriate to try if you would like.


I look forward to receiving your reply.



Thank y

Read More

Answer

I am very pleased to hear you would like to accept our offer and test these products in Flow Cytometry. Each code will give you: 1 free PRIMARY ANTIBODY OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for Flow Cytometry for the product and include the related code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. You do not need to referencea quote number when ordering these products. 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: www.abcam.com/collaborationdiscount.

Read More

Answer

Thank you for contacting us.

I have had a chance to review each of the product which you have listed as well as our other products against the same targets. I would like to offer some recommendations.

Anti GABA A receptor alpha 1 subunit antibody: ab33299 (https://www.abcam.com/GABA-A-Receptor-alpha-1-antibody-ab33299.html) and ab86320 (https://www.abcam.com/GABA-A-Receptor-alpha-1-antibody-ab86320.html) will both recognize the extracellular region of this protein in human tissue. Each will work as well as the other with the only difference being that ab86320 was created fromHuman GABA A Receptor alpha 1 andhas 100% homology, while ab33299 is created fromRat GABA A Receptor alpha 1with 93% homology.

Anti GABA A receptor beta 2 subunit antibody: Ab42598 (https://www.abcam.com/GABA-A-Receptor-beta-2-antibody-ab42598.html)is raised against the extracellular region of this protein and will react with human tissues.

Anti GABA B receptor 1 antibody: ab90883 (https://www.abcam.com/GABA-B-Receptor-1-antibody-ab90883.html) recognizes a cytoplasmic region of this protein and will require permeabilzation of the cells to be effective. Alternately ab55051 (https://www.abcam.com/GABA-B-Receptor-1-antibody-ab55051.html) recognizes the extracellular region of this protein. This will also work with human tissues.

None of our antibodies against these targets have been tested in flow cytometry. As such I may offer you enrollment in our 100% Abreview Testing Discount. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

I hope this information is helpful to you. Please let me know if you have any questions or would like to take advantage of our testing discount with these products.


Use our products? Submit an Abreview. Earn rewards!
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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Retina)
Specification
Retina
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Sep 12 2011

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