Overview

  • Product name
  • Description
    Guinea pig polyclonal to GABA
  • Host species
    Guinea pig
  • Specificity
    Recognizes GABA. Staining was blocked by preabsorbing with 100uM GABA conjugated to glutaraldehyde. 500uM of similar conjugations of glutamic acid, glutamate and taurine failed to block staining.
  • Tested applications
    Suitable for: IHC-P, Electron Microscopy, ICC/IF, IHC-FoFr, IHC-Fr, ELISAmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Chemical/ Small Molecule conjugated to KLH via glutaraldehyde.

  • General notes

    Without colchicine pretreatment well stained cell bodies are visible in the cerebral cortex, cerebrallar cortex, superior colliculus and some brainstem raphe. With colchicine pretreatment, additional cell body staining is present in the interpeduncular nucleus and the dorsal column nuclei.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer
    Constituent: Whole serum
  • Purity
    Whole antiserum
  • Primary antibody notes
    Without colchine pretreatment well stained cell bodies are visible in the cerebral cortex, cerebrallar cortex, superior colliculus and some brainstem raphe. With colchine pretreatment, additional cell body staining is present in the interpeduncular nucleus and the dorsal column nuclei.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab17413 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. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Electron Microscopy Use at an assay dependent concentration. PubMed: 20206640
ICC/IF 1/1000.
IHC-FoFr 1/1000.
IHC-Fr 1/1500.
ELISA Use at an assay dependent concentration.

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, labelling GABA with ab17413 diluted 1/500. Heat-induced antigen retrieval. DAB staining. Diffuse cytoplasmic staining and axonal staining observed in ventral midbrain neurons. (Note: This antibody produces diffuse cytoplasmic staining of moderate intensity in ventral midbrain, cortical and purkinje neurons in rat brain tissue). 

References

This product has been referenced in:
  • Chang JH  et al. Generation of Functional Dopaminergic Neurons from Reprogramming Fibroblasts by Nonviral-based Mesoporous Silica Nanoparticles. Sci Rep 8:11 (2018). Read more (PubMed: 29311646) »
  • Sullivan CS  et al. Perineuronal Net Protein Neurocan Inhibits NCAM/EphA3 Repellent Signaling in GABAergic Interneurons. Sci Rep 8:6143 (2018). Read more (PubMed: 29670169) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 13 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Cerebral Organoid)
Specification
Human Cerebral Organoid
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde

Radhika Menon

Verified customer

Submitted Sep 06 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Apteronotus leptorhynchus Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
Yes - 0.3% Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 09 2015

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (LDT brain slices)
Specification
LDT brain slices
Permeabilization
Yes - TritonX
Fixative
PFA/Gluteraldehyde

Jason Teem

Verified customer

Submitted Sep 10 2014

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Sample
Chicken Tissue sections (chicken hindbrain)
Specification
chicken hindbrain
Permeabilization
Yes - 0.1% PBS-TritonX
Fixative
Paraformaldehyde

Karli Montague

Verified customer

Submitted Mar 26 2014

Question
Answer


I can confirm that ab17413 anti-GABA antibody is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usually determined by protein assay, and serum will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for whole antiserum, concentration of antibody is known to very between 1 - 10 mg/ml.

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Answer

Thank you for bringing this to our attention. We have not tested the antibody on free-floating sections but your protocol is correct, assuming 10 minutes for permeabilization of these 50um sections was sufficient. We have one other complaint about this lot from a customer who was trying to stain similar samples. We do have a different lot now. If you like, I will be happy to send a vial as a free replacement, or a different antibody altogether, such as the rabbit polyclonal ab8891:

Click here (or use the following: https://www.abcam.com/index.html?datasheet=8891).

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Answer

Thank you for contacting us.

This antibody has been tested on GABAergic interneurons of mammalian samples, but we do not have testing or reports using hESC.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for providing some further useful information.

Both antibodies would be suitable for the immunostaining. However, it seems that ab17413 is more popular amongst our customers since it has been characterized in a number of applications. Moreover, we have one published feedback (see Abreview) and several specific published papers are available.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Answer

Thank you for contacting Abcam regarding ab17413. Regarding the tissue fixation, I have confirmed with the laboratory that yes, the combination of 4% paraformaldehyde / glutaraldehyde fixation is typically used in successful IHC staining.  Please find the protocol  is below: Immunohistochemistry Protocol: Fixation:  Tissues fixed with 4% paraformaldehyde and 0-0.5% glutaraldehyde give good results. 1. Tissues are fixed with 0.1M phosphate buffer, pH 6.5, 4% paraformaldehyde, 0-0.5% glutaraldehyde, 0.5% potassium dichromate. Tissue post-fixed overnight. 2. Cut sections in 50um thickness. 3. Incubate in 0.05M Tris buffer, pH 6.5 for 3 hrs. 4. Sections are incubated for up to 18-24 hours in ab17413 diluted with PBS, 0.1% sodium azide, 0.2% Triton X-100, 1% normal goat serum. 5. Fluorescein conjugated antibody or PAP may be used as the secondary reagent. I hope this information is helpful.  Please do not hesitate to contact me if you have any additional questions.

Read More

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,  

Read More
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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1-10 of 13 Abreviews or Q&A

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

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