MAB351 | Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6

Mouse anti-Glutamate Decarboxylase [GAD] (Catalog Number MAB351) localization of GAD in rat hippocampus. Photo courtesy of Dr. Robert Sloviter, University of Arizona.
Mouse anti-Glutamate Decarboxylase [GAD] (Catalog Number MAB351) localization of GAD in rat hippocam
MAB351
100 µg  
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      Overview

      Key Spec Table

      Species Reactivity Key Applications Host Format Antibody Type
      R,HWB,IHCMPurifiedMonoclonal Antibody
      Description
      Catalogue Number MAB351
      Brand Family Chemicon®
      Synonyms GAD65
      Trade Name
      • Chemicon
      Description Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6
      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. 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). GAD67 expression seems to be particularly plastic and can change in response to experimental manipulation (for example neuronal stimulation or transection) or disease progression and emergent disorders like schizophrenia (Volk et al., 2000). Colocalization of the two GAD isoforms also shows changes in GAD65/GAD67 distributions correlated with certain disease states such as IDDM and SMS.
      References
      Product Information
      Format Purified
      Specificity Recognizes the lower molecular weight isoform of the two GAD isoforms identified in brain (Gottlieb, et al., 1986; Chang & Gottlieb, 1988). This monclonal antibody can be used for immunohistochemical localization in brain or pancreas. Anti-GAD has also been used to label purified GAD on Western blots (Chang & Gottlieb, 1988).
      Control
      • human brain lysate
      • Rat brain tissue
      Presentation Purified immunoglobulin. Lyophilized from 10 mM potassium phosphate, 70 mM sodium chloride, pH 7.4, 0.02% sodium azide. Reconstitute with 100 μL of sterile water (1 mg/mL).
      Applications
      Application Anti-Glutamate Decarboxylase Antibody, 65 kDa isoform, clone GAD-6 detects level of Glutamate Decarboxylase & has been published & validated for use in IH & WB.
      Key Applications
      • Western Blotting
      • Immunohistochemistry
      Application Notes Immunohistochemistry: ( 1 μg/mL * See protocol)

      Western blot

      Optimal working dilutions must be determined by end user.

      APPLICATION NOTES FOR MAB351

      IMMUNOHISTOCHEMISTRY

      1) Perfuse rats with 100 mM phosphate buffer, pH 7.4 containing 1% paraformaldehyde, 0.34% L-lysine and 0.05% m-periodate (1% PLP).

      2) Postfix brains in 1% PLP for 1-2 hours.

      3) Transfer brains to 100 mM phosphate buffer containing 30% sucrose and gently agitate on a shaker platform at +4°C for 48-60 hours.

      4) Using a sliding microtome, cut 30 mm sections of frozen cerebellum. As the sections are cut, collect them in a vial of cold 100 mM phosphate buffer.

      5) Incubate sections in PBS containing 1.5% normal serum and 0.2% Triton X-100 for 30 minutes.

      6) On a shaker platform, incubate sections with MAB351 (diluted 1 μg/mL in PBS containing 1.5% normal serum and 0.2% Triton X-100) for 12-36 hours at +4°C.

      7) On a shaker platform, rinse sections eight times, 10-15 minutes per rinse, in PBS.

      8) Detect with standard secondary antibody detection system (PAP, ABC, etc.).

      9) Mount sections, dehydrate, and apply coverslips.
      Biological Information
      Immunogen Purified rat brain glutamic acid decarboxylase.
      Clone GAD-6
      Concentration Please refer to the Certificate of Analysis for the lot-specific concentration.
      Host Mouse
      Specificity Recognizes the lower molecular weight isoform of the two GAD isoforms identified in brain (Gottlieb, et al., 1986; Chang & Gottlieb, 1988). This monclonal antibody can be used for immunohistochemical localization in brain or pancreas. Anti-GAD has also been used to label purified GAD on Western blots (Chang & Gottlieb, 1988).
      Isotype IgG2a
      Species Reactivity Rat Human
      Antibody Type Monoclonal Antibody
      Entrez Gene Number
      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.
      Gene Symbol
      • EC 4.1.1.15
      • GAD2
      • MGC161607
      • GAD65
      • MGC161605
      • GAD-65
      Purification Method Protein A purfied
      UniProt Number
      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.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      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.
      Storage and Shipping Information
      Storage Conditions Maintain lyophilized material at -20°C for up to 12 months. After reconstitution maintain frozen at -20°C in undiluted aliquots for up to 6 months. Avoid repeated freeze/thaw cycles.
      Packaging Information
      Material Size 100 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Languages
      English

      Certificates of Analysis

      TitleLot Number
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE -2519307 2519307
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 1998916 1998916
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 2070808 2070808
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 2189668 2189668
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 2275543 2275543
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 2326979 2326979
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY - 2365949 2365949
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY -2475646 2475646

      References | 27 Available | See All References

      Reference overviewPub Med ID
      Reduced glutamate decarboxylase 65 protein within primary auditory cortex inhibitory boutons in schizophrenia.
      Caitlin E Moyer,Kristen M Delevich,Kenneth N Fish,Josephine K Asafu-Adjei,Allan R Sampson,Karl-Anton Dorph-Petersen,David A Lewis,Robert A Sweet
      Biological psychiatry 72 2012

      Show Abstract
      22624794
      Thalamocortical dysfunction and thalamic injury after asphyxial cardiac arrest in developing rats.
      Michael Shoykhet,Daniel J Simons,Henry Alexander,Christina Hosler,Patrick M Kochanek,Robert S B Clark
      The Journal of neuroscience : the official journal of the Society for Neuroscience 32 2012

      Show Abstract
      22492052
      Salubrinal, an endoplasmic reticulum stress blocker, modulates sleep homeostasis and activation of sleep- and wake-regulatory neurons.
      Methippara, M, et al.
      Neuroscience, 209: 108-18 (2012) 2012

      Show Abstract
      22387272
      Compensation of depleted neuronal subsets by new neurons in a local area of the adult olfactory bulb.
      Murata K, Imai M, Nakanishi S, Watanabe D, Pastan I, Kobayashi K, Nihira T, Mochizuki H, Yamada S, Mori K, Yamaguchi M.
      The Journal of neuroscience : the official journal of the Society for Neuroscience 31 10540-57 2011

      Show Abstract
      21775599
      Synaptic reorganisation of the medial amygdala during puberty.
      Cooke BM
      J Neuroendocrinol 23 65-73. doi 2011

      Show Abstract
      20874778
      Complementary distribution of glutamatergic cerebellar and GABAergic basal ganglia afferents to the rat motor thalamic nuclei.
      Eriko Kuramoto,Fumino Fujiyama,Kouichi C Nakamura,Yasuhiro Tanaka,Hiroyuki Hioki,Takeshi Kaneko
      The European journal of neuroscience 33 2011

      Show Abstract
      21073550
      Interneurons in the developing human neocortex.
      Nada Zecevic,Frances Hu,Igor Jakovcevski
      Developmental neurobiology 71 2011

      Show Abstract
      21154907
      Nitric oxide signaling modulates synaptic transmission during early postnatal development.
      Csaba Cserép,András Szonyi,Judit M Veres,Beáta Németh,Eszter Szabadits,Jan de Vente,Norbert Hájos,Tamás F Freund,Gábor Nyiri
      Cerebral cortex (New York, N.Y. : 1991) 21 2011

      Show Abstract Full Text Article
      21282319
      Evaluation of AAV-Mediated Expression of Chop2-GFP in the Marmoset Retina.
      Ivanova E, Hwang GS, Pan ZH, Troilo D
      Invest Ophthalmol Vis Sci 2010

      Show Abstract
      20484599
      Reduced cortical BDNF expression and aberrant memory in Carf knock-out mice.
      Kelli A McDowell,Ashley N Hutchinson,Sarah J E Wong-Goodrich,Matthew M Presby,Dan Su,Ramona M Rodriguiz,Krystal C Law,Christina L Williams,William C Wetsel,Anne E West
      The Journal of neuroscience : the official journal of the Society for Neuroscience 30 2010

      Show Abstract Full Text Article
      20519520

      Data Sheet

      Title
      MOUSE ANTI-GLUTAMIC ACID DECARBOXYLASE MONOCLONAL ANTIBODY

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      Categories

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