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Purified Mouse Anti-GAD65
Purified Mouse Anti-GAD65

Western blot analysis of GAD65 expression on rat brain. Lysates from rat brain cortex were probed with Purified Mouse Anti-GAD65 (Cat. No. 559931) at a concentration of 5.0, 2.0, or 0.5 µg/ml (lanes 1, 2, or 3 respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 554002). GAD65 is identified as a protein of ~65 kDA.  

Western blot analysis of GAD65 expression on rat brain. Lysates from rat brain cortex were probed with Purified Mouse Anti-GAD65 (Cat. No. 559931) at a concentration of 5.0, 2.0, or 0.5 µg/ml (lanes 1, 2, or 3 respectively), followed by HRP Goat Anti-Mouse Ig (Cat. No. 554002). GAD65 is identified as a protein of ~65 kDA.  

Product Details
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BD Pharmingen™
Glutamic acid decarboxylase
Rat (QC Testing), Human, Mouse, Pig (Tested in Development)
Mouse IgG2a
Purified Rat GAD65
Western blot (Routinely Tested), Immunohistochemistry-frozen, Immunoprecipitation (Reported)
65 kDa
0.5 mg/ml
AB_397380
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Rat brain cortex is recommended as a positive control for western blot

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
559931 Rev. 4
Antibody Details
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GAD-6

γ-aminobutyric acid (GABA) is an amino acid neurotransmitter that is considered to be the major inhibitory neurotransmitter in the mammalian central nervous system. The highest concentrations of GABA are found in the brain, where it is synthesized from glutamic acid to GABA by an enzyme called glutamic acid decarboxylase (GAD). GAD is also expressed in the insulin-producing β cells of the islets of Langerhans. Two isoforms of GAD are present in rat brain, GAD65 and GAD67, based on their relative molecular weight in kDa. Both isoforms have significant levels of homology in the catalytic portion of the molecule, but differ greatly in the first 95 amino acids in the N-terminal region. GAD65 migrates at ~65 kDa in SDS/PAGE. The antibody is reported to recognize rat, human, mouse, and pig GAD65. Purified GAD65 from rat brain was used as the immunogen.  The specific epitope recognized by this clone is a linear epitope localized in the last 41 amino acids of GAD65.

559931 Rev. 4
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
559931 Rev.4
Citations & References
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Development References (7)

  1. Bu DF, Erlander MG, Hitz BC. Two human glutamate decarboxylases, 65-kDa GAD and 67-kDa GAD, are each encoded by a single gene. Proc Natl Acad Sci U S A. 1992; 89(6):2115-2119. (Biology). View Reference
  2. Chang YC, Gottlieb DI. Characterization of the proteins purified with monoclonal antibodies to glutamic acid decarboxylase. J Neurosci. 1988; 8(6):2123-2130. (Immunogen: Western blot). View Reference
  3. Esclapez M, Tillakaratne NJ, Kaufman DL, Tobin AJ, Houser CR. Comparative localization of two forms of glutamic acid decarboxylase and their mRNAs in rat brain supports the concept of functional differences between the forms. J Neurosci. 1994; 14(3 Pt 2):1834-1855. (Biology). View Reference
  4. Hsu CC, Thomas C, Chen W. Role of synaptic vesicle proton gradient and protein phosphorylation on ATP-mediated activation of membrane-associated brain glutamate decarboxylase. J Biol Chem. 1999; 274(34):24366-24371. (Clone-specific: Immunoprecipitation, Western blot). View Reference
  5. Kanaani J, Lissin D, Kash SF, Baekkeskov S. The hydrophilic isoform of glutamate decarboxylase, GAD67, is targeted to membranes and nerve terminals independent of dimerization with the hydrophobic membrane-anchored isoform, GAD65. J Biol Chem. 1999; 274(52):37200-37209. (Biology). View Reference
  6. Kaufman DL, McGinnis JF, Krieger NR, Tobin AJ. Brain glutamate decarboxylase cloned in lambda gt-11: fusion protein produces gamma-aminobutyric acid. Science. 1986; 232(4754):1138-1140. (Biology). View Reference
  7. Kim J, Richter W, Aanstoot HJ . Differential expression of GAD65 and GAD67 in human, rat, and mouse pancreatic islets. Diabetes. 1993; 42(12):1799-1808. (Clone-specific: Immunohistochemistry, Western blot). View Reference
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559931 Rev. 4

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.