Anti-Glutamate Decarboxylase 65 & 67 Antibody

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.

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.

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

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

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

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.

65/68 kDa

Ammonium Sulfate Precipitation

Format: Purified

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

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.

Control
Whole rat brain lysate, mouse brain cell extract.

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

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