GAD67 Antibody Staining Protocol for Immunohistochemistry

Description: Glutamic acid decarboxylase (GAD) catalyses the conversion of L-glutamic acid to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Two forms of human GAD, GAD65 and GAD67, are encoded by two separate genes. Human GAD65 cDNA encodes a Mr 65,000 polypeptide, with 585 amino acid residues, whereas human GAD67 encodes a Mr 67,000 polypeptide, with 594 amino acid residues. GAD67 gene consists of 16 exons, spread over more than 45 kb of genomic DNA. The GAD67 gene contains an additional exon (exon 0) that together with part of exon 1, specifies the 5' untranslated region of GAD67 mRNA. Human GAD67 shows 65% identity to GAD65 and is located in 2q31. GAD67 may play a role in the stiff man syndrome. Deficiency in this enzyme has been shown to lead to pyridoxine dependency with seizures.

Primary Antibody

Name: GAD67 IHC Antibody

Clone: Rabbit polyclonal

Supplier: IHC World

Catalog Number: IW-PA1036

Dilution: Ready to Use

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval

Device: IHC-Tek^TM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-Tek^TM Epitope Retrieval Solution (Cat# IW-1100)

Heat/Cool Temperature: 95-100 ºC/room temperature

Heat/Cool Time: 20 minutes/20 minutes

Detection Methods

Standard Method: ABC Method or LSAB Method

Enhanced Method: Polymeric Methods

Chromogen Substrate

Reagent: DAB

Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Cytoplasmic

Images: Search image

Additional Information:

Species Reactivity: Human, mouse, rat

Fixation: Formalin fixed paraffin sections

Positive Control: Brain

Negative Control: Omit primary antibody, isotype control, absorption control

Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

References: