Glutamic acid decarboxylase polyclonal antibody

Synthetic peptide corresponding to aa 572-585 (D572FLIEEIERLGQDL585) of rat glutamate decarboxylase (GAD65) . This sequence is conserved in rat GAD67 and in human GAD-2.

A synthetic peptide with the amino acid sequence: [C]-D-F-L-I-E-E-I-E-R-L-G-Q-D-L was selected from rat glutamate decarboxylase (GAD65; C-terminus residues [Cys]+572-585) . This sequence is conserved in rat GAD67 and in human GAD-28. The peptide was conjugated to keyhole limpet hemocyanin via the N-terminal cysteine residue using m-maleimidobenzoic acid N-hydroxysuccinimide. The conjugate was used to immunise New Zealand White rabbits and the resultant antiserum (B4; Z00257) has been extensively characterised by ELISA, Western blot and immunohistochemical techniques. Application data Western blotting Test tissues: Mouse and rat brain. Dilution: 1:1000. Protein size: Doublet on SDS-PAGE at approximately 65/67kDa. Immunohistochemistry Test tissues: Human, cat and rat brain (esp. cerebellum) . Fixatives: Several fixative solutions may be used. Aldehyde-combination fixatives (i.e. those containing formaldehyde and glutaraldehyde) usually give satisfactory results. Bouin and Susa fixatives containing 0.1-0.2% glutaraldehyde have been used satisfactorily. Processing: Cryostat and Vibratome®9 sections have been used most commonly. Paraffin wax-embedded tissue has been used successfully. The antiserum has also been used for the ultrastructural detection of GAD using pre-embedding immunocytochemistry. Dilution range: 1:200-1:5000, using overnight incubation and PAP or ABC-peroxidase procedure.