Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-HSP27 (Ser82) Antibody #2401

Applications Reactivity MW (kDa) Source
W IHC-P IHC-F IF-IC H Mk 27 Rabbit

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-HSP27 (Ser82) Antibody detects endogenous HSP27 only when phosphorylated at serine 82. The antibody does not recognize other heat shock proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho- peptide (KLH coupled) corresponding to residues surrounding Ser82 of human HSP27. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, using Phospho-HSP27 (Ser82) Antibody (upper) or control HSP27 antibody (lower). HeLa cells were incubated at 42°C for 0-2 hours as indicated.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (A,C) or lambda phosphatase-treated (B,D), using Phospho-HSP27 (Ser82) Antibody (A,B) or HSP27 (G31) Monoclonal Antibody #2402 (C,D).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-HSP-27 (Ser82) Antibody in the presence of contol peptide (left) or antigen specific peptide (right).


IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, showing cytoplasmic localization using Phospho-HSP27 (Ser82) Rabbit mAb.

IC-ABC

IC-ABC

Immunocytochemical staining of HeLa cells, untreated (upper) or UV-treated (lower), using anti-Phospho-HSP27 (Ser82) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, anisomycin-treated (left) or untreated (right), using Phospho-HSP27 (Ser82) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


Background

Heat shock protein (HSP) 27 is one of the small HSPs that are constitutively expressed at different levels in various cell types and tissues. Like other small heat shock proteins, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78 and Ser82 by MAPKAP kinase 2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6).

  1. Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
  2. Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
  3. Rouse, J. et al. (1994) Cell 78, 1027-1037.
  4. Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
  5. Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
  6. Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.

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