Overview

  • Product name
  • Description
    Rabbit polyclonal to GABA
  • Host species
    Rabbit
  • Specificity
    ab8891 targets conjugated GABA (Gamma-Aminobutyric acid), it does not recognize free GABA.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IHC-FoFrmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to GABA. Synthetic GABA conjugated to bovine serum albumin.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Constituents: ddH20, 50% Glycerol
  • Purity
    Whole antiserum
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8891 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 1/1000 - 1/5000. Fixation of tissue for use with these antibodies should be done with glutaraldehyde. The use of paraformaldehyde in conjunction with glutaraldehyde may improve staining results.
ICC/IF Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration. PubMed: 17394161

Target

  • Relevance
    Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter. GABA acts at inhibitory synapses in the brain and spinal cord. Inhibition is provoked by GABA binding resulting in hyperpolarization of the synaptic transmembrane potential of the affected neuron. GABA binding causes ion channels to open allowing either the flow of chloride or potassium ions into or out of the cell.
  • Cellular localization
    Cell Membrane, Cytoplasmic, Plasma membrane and Secreted
  • Alternative names
    • 4 aminobutanoic acid antibody
    • GABA antibody
    • Gamma amino butyric acid antibody
    • Gamma aminobutyric acid antibody

Images

  • Immunohistochemical analysis of rat brain tissue, staining GABA with ab8891.

    Slices were fixed in a 2% glutaraldehyde, 2% formaldehyde solution before being blocked for 2 h at room temperature in PBS with 0.3% Tween-20, 0.2% BSA and 5% normal goat serum. Samples were incubated with primary antibody (1/200) before detection with a fluorescence-conjugated goat anti-rabbit secondary antibody.
  • Immunofluorescent analysis of Human embryonic stem cells, staining GABA with ab8891.

    Cells were fixed with paraformaldehyde before blocking with 10% goat serum, 1% BSA, 0.1% Triton X-100 for 1 hour. Samples were stained with primary antibody (1/500) overnight at 4°C. An AlexaFluor®594-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.

References

This product has been referenced in:
  • Zieger E  et al. Roles of Retinoic Acid Signaling in Shaping the Neuronal Architecture of the Developing Amphioxus Nervous System. Mol Neurobiol N/A:N/A (2017). Read more (PubMed: 28875454) »
  • Ogundele OM  et al. Systemic Sympathoexcitation Was Associated with Paraventricular Hypothalamic Phosphorylation of Synaptic CaMKIIa and MAPK/ErK. Front Neurosci 11:447 (2017). Read more (PubMed: 28824368) »
See all 10 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

The optimal incubation time will depend on how your tissue sections are prepared.

The protocol used for the stain of rat brain tissue shown on the datasheet is from this reference:

Margolis EB et al. Identification of rat ventral tegmental area GABAergic neurons. PLoS One 7:e42365 (2012).

Here is a link: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3409171/

They were working with thick vibratome sections of 150-200 um, so they incubated them for 48 hours at 4C in a 1/200 dilution of the antibody. If your sections are thin, for instance 5 - 10 um, incubation overnight at 4C should be sufficient.

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 22°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (early LDT coronal sections)
Specification
early LDT coronal sections
Permeabilization
Yes - TritonX-100
Fixative
Paraformaldehyde

Jason Teem

Verified customer

Submitted Jan 08 2014

Answer

Thank you for contacting us.
The antibody has been tested in western blot in conjunction with the antigen as a positive control, in which case the band would is ˜67 kDa as the antigen is made with BSA. Free GABA will not be an option. Whatever is loaded onto the blot will need to have a gluteraldehyde component included somewhere in the prep of the samples to complete the epitope for the antibody.

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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Question

Further to our below email exchange concerning the ab8891 antibody, please find hereby the reply from the customer to your questionnaire: 1.       Order Details Antibody code: ab8891 2.       Lot number: GR-46247-1 3.       General Information Antibody storage conditions (temperature/reconstitution etc) : 4C 4.       Description of the problem (high background, low signal, non-specific staining etc.) : no signal 5.       Sample (Species/Tissue/Cell Type/Cell Line etc.) mouse frozen brain sections, embryonic and adult. 6.       Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) 4% PFA (O/N) and 4%PFA (O/N) + 0.25% glutaraldehyde (10 minutes) 7.       Antigen retrieval (Enzymatic method, Heat mediated technique etc.) : none 8.       Permeabilization step : 0.3% Triton X 9.       Blocking conditions (Buffer/time period, Blocking agent etc.) : 1% bovine serum albumin, 5% goat serum and 0.3% Triton-X in 1x PBS for at least 1h at RT 10.   Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) tried 1-200 to 1:2000, O/N at room temperature. 11.   Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Which secondary was used? Is this working well with other primary antibodies? Invitrogen-Molecular probes Alexa fluor dyes, work great with other antibodies. 12.   Detection method : fluorescence 13.   Positive and negative controls used (please specify) : yes, no primary Ab (negative control) and embryonic tissue with other antibodies such as TH (ab112) (positive control) 14.   Optimization attempts (problem solving) How many times have you tried the IHC? 4 times 15.   Have you run a "No Primary" control? Yes 16.   Do you obtain the same results every time? Yes 17.   What steps have you altered? Fixation, dilution of primary, type of tissue 18.   Additional Notes :   I trust this information is to your satisfaction. Should you need any further inquiries please don’t hesitate to contact me.   Best regards,  

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality. Reviewing this case, I would like to offer some suggestions to help optimize the results from ab8891. I would also appreciate if you can confirm some further details:   1. I can recommend to try antigen retrieval as the tissue has been in PFA overnight, which is quite a long time. 2. Could you provide further details of the permebilization? I can suggest to try 0.1% Triton for 10 minutes only as Triton is quite a strong detergent, and can disrupt protein structure. 3. I can suggest to Use 0.2% Tween (a more gentle detergent) in the antibody dilution buffer, wash buffer and blocking buffer. 4. We recommend not to mix blocking agents in an experiment. Try 5% goat serum only. In the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.  

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Question
Answer

Thank you for your enquiry. According to the reference available for ab8891, they injected rabbits with a GABA-glutaraldehyde-lysine conjugate. I am still awaiting further information regarding ab17390. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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Answer

The originator of this antibody has informed me that they do not know of any species ab8891 does NOT work in successfully. So, it should work in rat. Please let me know if your customer has any more questions.

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Answer

This antibody has been tested and characterised for ELISA application. All the information available for this product is listed on the on-line datasheet (price, datasheet, publication, suitability, cross-reactivity). I would suggest that perhaps you should have a look at the published references listed on this product datasheet, you may find some useful information there.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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