DARPP 32 (Phospho-Phospho-Thr34) Antibody

DARPP 32 (Phospho-Phospho-Thr34) Antibody | catalog# XPS-1004

Background

DARPP-32, a dopamine (DA) and cAMP-regulated ~32k phosphoprotein that is associated with dopaminoceptive neurons bearing D-1 receptors in the basal ganglia. The protein inhibits protein phosphatase I when it is phosphorylated on Thr34. In contrast, when DARPP-32 is phosphorylated on Thr75 the protein acts as an inhibitor of PKA. Phosphorylation of DARPP-32 is thought to play a critical role in the regulation of dopaminergic neurotransmission. In addition, the activity of DARPP-32 is also thought to play important roles in the actions of alcohol, caffeine and Prozac®.

Description

DARPP 32 (Phospho-Phospho-Thr34) Antibody

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Specifications

Clonality/Clone

This is a polyclonal antibody.

Purification

Affinity Purified

Host

DARPP 32 antibody was raised in rabbit.

Please use anti-rabbit secondary antibodies.

Immunogen

Rabbit anti-DARPP-32 (Thr34) polyclonal antibody was raised against a synthetic phosphopeptide corresponding to amino acids residues surrounding the phospho Thr34 of DARPP-32. Rabbit anti-DARPP-32 (Thr34) was purified by sequential chromatography on phospho-and non-phosphopeptide affinity columns.

Application

DARPP 32 antibody is specific for the ~32k DARPP-32 phosphorylated at Thr34. Applications include Dot Blots (DB) and Western Blots (WB). Suitability for Immunohistochemistry (IHC) has not yet been determined. Rabbit anti-DARPP-32 (Thr34) recognizes human, mouse, and rat forms of the protein. When internally tested under ideal conditions the working dilutions were 1:1000 for DB and WB.

Tested Application

Buffer

DARPP 32 antibody is supplied as affinity purified polyclonal antibody, 100 μl in 10 mM HEPES (pH 7.5), 150 mM NaCl, 100 μg/ml BSA and 50% glycerol, and there is adequate amount of material to conduct 10-mini Western Blots.

Storage

For long term storage -80˚C is recommended, but shorter term storage at -20˚C is also acceptable as aliquots may be taken without freeze/thawing due to the presence of 50% glycerol. Stock solutions are stable for a minimum of 1 year at -20˚C.

Species Reactivity

H, M, R, X

Reference

Svenningsson, P., Tzavara, E.T., Carruthers, R., Rachleff, I., Wattler, S., Nehls, M., McKinzie, D.L., Fienberg, A.A., Nomikos, G.G.and Greengard, P., “Diverse psychotomimetics act through a common signaling pathway,” Science, 302 (2003) 1412 - 1415.
Lindskog, M., Svenningsson, P., Pozzi, L., Kim, Y., Fienberg, A.A., Bibb, J.A.., Fredholm, B.B., Nairn, A.C., Greengard, P. and Fisone, G., “Involvement of DARPP-32 phosphorylation in the stimulant action of caffeine, Nature (London), 418 (2002) 774 - 778.
Maldve, R.E., Zhang, T.A., Ferrani-Kile, K., Schreiber, S.S., Lippmann, M.J., Snyder, G.L., Feinberg, A.A., Leslie, S.W., Gonzales, R.A., and Morrisett, R.A., “DARPP-32 and the regulation of the ethanol sensitivity of NMDA receptors in the nucleus accumbens,” Nature Neurosci. 5 (2002) 641 - 648.
Bibb, J.A., Snyder, G.L., Nishi, A., Yan, Z., Meijer, L., Fienberg, A.A., Tsai, L.H., Kwon, Y.T., Girault, J.A., Czernik, A.J., Huganir, R.L., Hemmings, Jr., H.C., Nairn, A.C., and Greengard, P., “Phosphorylation of DARPP-32 by Cdk5 modulates dopamine signalling in neurons, Nature (London) 402 (1999) 669 - 671.]