Antigen/Purification: Collapse
The antigen is a fusion protein from the cytoplasmic loop of the α6-subunit of rat GABAA receptor.
The antibody is prepared from rabbit serum by affinity purification using a column to which the fusion protein immunogen was coupled.
Biological Significance: Collapse
Gamma-aminobutyric acid (GABA) is the primary inhibitory neurotransmitter in the central nervous system, causing a hyperpolarization of the membrane through the opening of a Cl− channel associated with the GABAA receptor (GABAA-R) subtype. GABAA-Rs are important therapeutic targets for a range of sedative, anxiolytic, and hypnotic agents and are implicated in several diseases including epilepsy, anxiety, depression, and substance abuse. The GABAA-R is a multimeric subunit complex. To date six αs, four βs and four γs, plus alternative splicing variants of some of these subunits, have been identified (Olsen and Tobin, 1990; Whiting et al., 1999; Ogris et al., 2004). Injection in oocytes or mammalian cell lines of cRNA coding for α- and β-subunits results in the expression of functional GABAA-Rs sensitive to GABA. However, coexpression of a γ-subunit is required for benzodiazepine modulation. The various effects of the benzodiazepines in brain may also be mediated via different α- subunits of the receptor (McKernan et al., 2000; Mehta and Ticku, 1998; Ogris et al., 2004; Pöltl et al., 2003).
Storage
100 µl in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 µg per ml BSA and 50% glycerol. Adequate amount of material to conduct 10-mini Western Blots.
For long term storage –20° C is recommended. Stable at –20° C for at least 1 year.
General References
McKernan RM, et al. (2000) Sedative but not anxiolytic properties of benzodiazepines are mediated by the GABAA receptor α1-subtype. Nature Neurosci 3:587-592.
Mehta AK, Ticku MK (1998) Prevalence of the GABAA receptor assemblies containing α1-subunit in the rat cerebellum and cerebral cortex as determined by immunoprecipitation: Lack of modulation by chronic ethanol administration. Mol Brain Res 67:194-199.
Ogris W, Pöltl A, Hauer B, Ernst M, Oberto A, Wulff P, Höger H, Wisden W, Sieghart W (2004) Affinity of various benzodiazepine site ligands in mice with a point mutation in the GABAA receptor γ2-subunit. Biochem Pharmacol 68:1621-1629.
Olsen RW, Tobin AJ (1990) Molecular biology of GABAA receptors. FASEB 4:1469-1480.
Pöltl A, Hauer B, Fuchs K, Tretter V, Sieghart W (2003) Subunit composition and quantitative importance of GABAA receptor subtypes in the cerebellum of mouse and rat. J Neurochem 87:1444-1455.
Whiting PJ, Bonnert TP, McKernan RM, Farrar S, Le Bourdellès B, Heavens RP, Smith DW, Hewson L, Rigby MR, Sirinathsinghji DJS, Thompson SA, Wafford KA (1999) Molecular and functional diversity of the expanding GABAA receptor gene family. Ann NY Acad Sci 868:645-653.
