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Structure-functions of HspB1 (Hsp27).

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

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Cytochalasin D, from Zygosporium mansonii, ≥98% (TLC and HPLC), powder
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Millipore
Protein G–Agarose, lyophilized powder, Contains lactose stabilizers that must be removed prior to use.