Glutamate Decarboxylase 65,67 (GAD 65,67)

Glutamic 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 (65kD and 67kD, 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 (585aa) encoded on human chromosome10. GAD67 is cytoplasmic (594aa) and is encoded on chromosome 2. GAD expression changes during neural development in rat spinal cord. GAD65 is expressed transiently in commissural axons around E13 but is down regulated the next day while GAD67 expression increases mostly in the somata of those neurons (Phelps, P. et al. 1999). In mature rat pancreas, GAD65 and GAD67 appear to be differentially localized, GAD65 primarily in insulin-containing beta cells and GAD67 in glucagon-containing (A) cells (Li, L. et al. 1995). Changes in GAD65/GAD67 distributions have also been correlated with certain disease states such as IDDM and SMS.\n\nApplications:\nSuitable for use in Immunohistochemistry, ELISA, Western Blot and Immunofluorescence. Other applications have not been tested.\n\nRecommended Dilutions:\nImmunohistochemistry (Frozen): 1:200-1:5000\nImmunohistochemistry: Paraffin sections\nWestern Blot: 1:1000-1:10,000. A doublet at ~65/67kD is expected.\nOptimal working dilutions to be determined by researcher.\n\nRecommended Positive Control Tissue: \nBrain\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

G4000-11

Size

100ul

Applications

ELISA, IF, IHC, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Rat, Fel

Form

Supplied as a liquid in PBS, 1% BSA, 0.09% sodium azide.

P Type

Pab

Purity

Purified by ammonium sulfate precipitation.

Isotype

IgG

References

US Biological application reference: Hughes, S.M. et al., (2009) Dev Neurobiol. 69:477-490. 1. Hernandez-Montiel, H.L. et al. (2003) Diffusible signals and fasciculated growth in reticulospinal axon pathfinding in the hindbrain. Dev. Biol. 255: 99-112.

Additional Info

Recognizes both molecular forms of rat glutamic acid decarboxylase \n(GAD65 and GAD67), the enzyme which converts glutamic acid to gamma-aminobutyric acid (GABA), the major inhibitory transmitter in the higher brain region. Species Crossreactivity: human, mouse and feline