Technical Data
G7116-05
Glutamic Acid Decarboxylase (GAD)
Description:
The enzyme glutamic acid decarboxylase (GAD) catalyzes the synthesis of g-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the central nervous system. GAD is the rate-limiting enzyme in the biosynthesis of GABA from L-glutamic acid, and it is used extensively as a marker for GABAergic neurons and synaptic terminals. GAD has been observed in pancreatic islet cells and identified as an autoantigen associated with the development of insulin-dependent diabetes mellitus.

Applications:
Suitable for use in Western Blot and Immunohistochemistry. Other applications not tested.

Recommended Dilution:
Immunohistochemistry: 1ug/ml
Optimal dilutions to be determined by the researcher.

Immunohistochemistry Protocol:
1. Perfuse rats with 100mM 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 100mM phosphate buffer containing 30% sucrose and gently agitate on a shaker platform at 4C for 48-60 hours.
4. Using a sliding microtome, cut 30um sections of frozen cerebellum. As the sections are cut, collect them in a vial of cold 100mM 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 G7116-05
(diluted 1ug/ml in PBS containing 1.5% normal serum and 0.2% Triton X-100) for 12-36 hours at 4C.
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.

Storage and Stability:
Lyophilized powder may be stored at 4C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol, ddH2O and store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG2a2Q61Purified
SizeStorageShippingSourceHost
100ug4C (-20C Glycerol)Blue IceRatMouse
Concentration:
~1mg/ml (after reconstitution)
Immunogen:
Purified rat brain glutamic acid decarboxylase (GAD).
Purity:
Purified
Form
Supplied as a lyophilized powder from PBS, pH 7.4. Reconstitute with 100ul sterile 40-50% glycerol, ddH2O.
Specificity:
Recognizes the lower MW isoform (65kDa) of the two GAD isoforms identified in brain. This monclonal antibody can be used for immunohistochemical localization in brain or pancreas. Has been used to label purified GAD on Western blots. Species Crossreactivity: Rat
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. PNAS USA 83: 88088812 (1986). 2. J. Neuroscience 8: 2123 (1988). 3. Liu, S., et al., J. Neuroscience 23: in press (2003). 4. Atkinson, L., et al., Neuroscience 123: 761-768 (2004).