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AB1511 Sigma-Aldrich

Anti-Glutamate Decarboxylase 65 & 67 Antibody

This Anti-Glutamate Decarboxylase 65 & 67 Antibody is validated for use in IH, WB for the detection of Glutamate Decarboxylase 65 & 67.

Synonym: 65 kDa glutamic acid decarboxylase, Glutamate decarboxylase 65 kDa isoform, Glutamate decarboxylase-2 (pancreas), glutamate decarboxylase 2, glutamate decarboxylase 2 (pancreatic islets and brain,65kD), glutamate decarboxylase 2 (pancreatic islets and br

  •  eCl@ss 32160702

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Properties

Related Categories Alphabetical Index, Antibodies, GD-GL, Primary Antibodies
clone   polyclonal
biological source   rabbit
application(s)   immunohistochemistry: suitable
  western blot: suitable
species reactivity   feline, mouse, human, rat
shipped in   dry ice
antibody product type   primary antibodies
mfr. no.   Chemicon®
NCBI accession no.   NP_000809
UniProt accession no.   Q05329
Gene Information   human ... GAD2(2572)

Description

General description

Gutamic acid decarboxylase (GAD; E.C. 4.1.1.15) is the enzyme responsible for the conversion of glutamic acid to gamma-aminobutyric acid (GABA), the major inhibitory transmitter in higher brain regions, and putative paracrine hormone in pancreatic islets. Two molecular forms of GAD (65 kDa and 67 kDa, 64% aa identity between forms) are highly conserved and both forms are expressed in the CNS, pancreatic islet cells, testis, oviduct and ovary. The isoforms are regionally distributed cytoplasmically in the brains of rats and mice (Sheikh, S. et al. 1999). GAD65 is an ampiphilic, membrane-anchored protein (585 a.a.), encoded on human chromosome 10, and is responsible for vesicular GABA production. GAD67 is cytoplasmic (594 a.a.), encoded on chromosome 2, and seems to be responsible for significant cytoplasmic GABA production. GAD expression changes during neural development in rat spinal cord. Colocalization of the two GAD isoforms also shows changes in GAD65/GAD67 distributions correlated with certain disease states such as IDDM and SMS.

Specificity

Glutamate Decarboxylase (GAD65 and GAD67).
Western blot reveals a doublet at approximately 65/68 kDa.
Immunohistochemical staining can be abolished by preincubation with 1-10 μg peptide (Catalog Number AG252) per mL of diluted antibody.

Immunogen

A synthetic peptide with the amino acid sequence [C]DFLIEEIERLGQDL from rat glutamate decarboxylase (GAD65; C-terminus residues [Cys] + 572-585) (1).

Application

Immunohistochemistry:
1:200-1:5,000 dilution of a previous lot was used in immunohistochemistry (overnight PAP or ABC). Several fixative solutions may be used. Aldehyde-combination fixatives (i.e. those containing formaldehyde and glutaraldehyde) usually give satisfactory results. Bouin and Susa fixatives containing 0.1-0.2% glutaraldehyde have been used satisfactorily. Also reactive on paraffin embedded tissue. Protease digestion is not required.

Optimal working dilutions must be determined by the end user.

Research Category
Neuroscience

Research Sub Category
Neurotransmitters & Receptors

Neuronal & Glial Markers

Target description

65/68 kDa

Physical form

Ammonium Sulfate Precipitation

Format: Purified

Purified rabbit polyclonal in buffer containing PBS-BSA containing 0.1% sodium azide.

Storage and Stability

Stable for 1 year at -20°C in undiluted aliquots (not less than 10 μL) from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Quality

Evaluated by Western Blot on mouse brain lysates.

Western Blot Analysis:
1:500 dilution of this antibody detected GAD 65&67 on 10 μg of mouse brain lysates.

Analysis Note

Control
Whole rat brain lysate, mouse brain cell extract.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

Please Enter a Lot Number
Protocols & Articles

Articles

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detection methods, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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