Structure-functions of HspB1 (Hsp27).

Methods in molecular biology (Clifton, N.J.) (2011-09-08)
André-Patrick Arrigo

Human HspB1 (also denoted Hsp27) is a well-known member, together with alphaB-crystallin, of the small heat-shock (or stress) proteins (sHsps) (20-40 kDa). In this chapter, I describe procedures for testing the oligomeric and phosphorylation patterns of HspB1 as well as its interaction with specific partner/client polypeptides using tissue culture cells genetically modified to express different levels of this protein. The procedures have been developed in my laboratory and could be used in any well-established cellular laboratory. In addition, the different procedures presented here could be extended to test the nine other human sHsp members as well as sHsps from other species.

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Protein G–Agarose, lyophilized powder, Contains lactose stabilizers that must be removed prior to use.