Human Hsp27 belongs to the phylogenically conserved small heat shock protein (smHsp) family, which includes mouse Hsp25, alphaA-crystallins and alphaB-crystallins.
Members of this protein superfamily from the animal, plant and microbiotic kingdoms share the so-called alpha-crystallin domain of approximately 90 amino acid residues which is bounded by variable N-and C-terminal extensions (1).
Hsp27 is expressed constitutively in many tissues and its expression is increased to high levels after various types of stress including elevated temperatures, toxic metals, drugs and oxidants.
Hsp27 is believed to exist mainly as oligomers of as many as 8–40 of Hsp27 protein monomers in cells and data suggest that these large oligomers have a chaperone-like activity by serving as a site where unfolding proteins may bind until ATP and Hsp70-dependent refolding can occur (2).
Hsp27 is phosphorylated on three phosphorylation sites (Ser15, Ser78 and Ser82) by protein kinases including MAPKAP kinase 2/3 and the stress-activated protein kinase SAPK2 (p38) (3,4).
Studies on cells stimulated by a variety of mitogenic and stress factors suggest that phosphorylation of Hsp27 occurs as an early prominent event and that phosphorylation induced changes in the ultrastructure of Hsp27 may regulate its biochemical activities.
The state of phosphorylation and oligomerization of Hsp27 may regulate microfilament organization as shown by studies demonstrating that only the nonphosphorylated lower molecular weight forms of Hsp27 bind actin barbed ends and inhibit polymerization (5).
Hsp27 is believed to protect cells by enhancing cellular glutathione levels as indicated by elevated glutathione levels in cells overexpressing Hsp27.
Studies using wild-type Hsp27 and mutants in which the serine phosphorylation sites were mutated to alanines, glycines or aspartates, demonstrate that cellular glutathione levels depend on the oligomerization of Hsp27 with only the large oligomeric forms of Hsp27 capable of protecting cells by enhancing glutathione levels (6).
Applications: Suitable for use in Western Blot and Immunohistochemistry.
Other applications have not been tested.
Recommended Dilution:Western Blot (ECL): 1:1000Immunohistochemistry: 1:50Optimal dilutions to be determined by researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：50ug
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by peptide affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.2, 0.09% sodium azide, 50% glycerol.
Specificity：Recognizes human phosphorylated Hsp27 (Ser15) at ~27kD, corresponding to the MR of phosphorylated Hsp27 (Ser15) on SDS-PAGE immunoblots. Does not crossreact with non-phosphorylated Hsp27. Species Crossreactivity: mouse, rat, bovine, canine, guinea pig and monkey.
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Bo Ca Gp Hu Mk
Immunogen：Synthetic peptide corresponding to a portion of human HSP27 phosphorylated at Ser15.
