Product Images
Rabbit anti-GAD65/67 (Catalog Number AB1511) staining of GAD65/67 in rat cerebellum. Photo courtesy of Dr. Stylianopoilou, University of Athens.
Western blot luminograph of whole rat brain lysate immuno-stained using the rabbit polyclonal antiserum to GAD65/67 showing both isoforms. Antiserum t...
see more >)
)
)
)
Anti-Glutamate Decarboxylase 65 & 67
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R, M, Fe | IH, WB | Rabbit | Purified | Polyclonal Antibody |
Description:
Anti-Glutamate Decarboxylase 65 & 67
Trade Name:
Chemicon (Millipore)
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.
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.
Molecular Weight:
65/68 kDa
Immunogen:
A synthetic peptide with the amino acid sequence [C]DFLIEEIERLGQDL from rat glutamate decarboxylase (GAD65; C-terminus residues [Cys] + 572-585) (1).
Background Information:
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.
Species Reactivity:
- Human
- Rat
- Mouse
- Feline
Species Reactivity Note:
Human, mouse, rat, and feline.
Application Notes:
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.
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.
Control:
Whole rat brain lysate, mouse brain cell extract.
Quality Assurance:
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.
Western Blot Analysis:
1:500 dilution of this antibody detected GAD 65&67 on 10 μg of mouse brain lysates.
Purification Method:
Ammonium sulfate precipitation
Presentation:
Purified rabbit polyclonal in buffer containing PBS-BSA containing 0.1% sodium azide.
Storage Conditions:
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.
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.
UniProt Number:
Entrez Gene Number:
Gene Symbol:
- GAD2
- MGC161607
- GAD65
- MGC161605
- GAD-65
Alternate Names:
- 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 brain, 65kDa)
Usage Statement:
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.
Key Applications:
- Immunohistochemistry
- Western Blotting
Entrez Gene Summary:
This gene encodes one of several forms of glutamic acid decarboxylase, identified as a major autoantigen in insulin-dependent diabetes. The enzyme encoded is responsible for catalyzing the production of gamma-aminobutyric acid from L-glutamic acid. A pathogenic role for this enzyme has been identified in the human pancreas since it has been identified as an autoantibody and an autoreactive T cell target in insulin-dependent diabetes. This gene may also play a role in the stiff man syndrome.
UniProt Summary:
FUNCTION: SwissProt: Q05329 # Catalyzes the production of GABA.
COFACTOR: Pyridoxal phosphate.
SIZE: 585 amino acids; 65411 Da
SUBUNIT: Homodimer (By similarity).
SUBCELLULAR LOCATION: Cytoplasm, cytosol. Cytoplasmic vesicle. Cell junction, synapse, presynaptic cell membrane; Lipid-anchor. Golgi apparatus membrane; Peripheral membrane protein; Cytoplasmic side. Note=Associated to cytoplasmic vesicles. In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters.
PTM: Phosphorylated; which does not affect kinetic parameters or subcellular location. & Palmitoylated; which is required for presynaptic clustering.
SIMILARITY: SwissProt: Q05329 ## Belongs to the group II decarboxylase family.
COFACTOR: Pyridoxal phosphate.
SIZE: 585 amino acids; 65411 Da
SUBUNIT: Homodimer (By similarity).
SUBCELLULAR LOCATION: Cytoplasm, cytosol. Cytoplasmic vesicle. Cell junction, synapse, presynaptic cell membrane; Lipid-anchor. Golgi apparatus membrane; Peripheral membrane protein; Cytoplasmic side. Note=Associated to cytoplasmic vesicles. In neurons, cytosolic leaflet of Golgi membranes and presynaptic clusters.
PTM: Phosphorylated; which does not affect kinetic parameters or subcellular location. & Palmitoylated; which is required for presynaptic clustering.
SIMILARITY: SwissProt: Q05329 ## Belongs to the group II decarboxylase family.
Brand Family:
Chemicon
Product Name:
Anti-Glutamate Decarboxylase 65 & 67
Concentration:
Variable
Antibody Type:
Polyclonal Antibody
Qty/Pk:
50 µL
Format:
Purified
Host:
Rabbit