Product Pathways - MAPK Signaling
Phospho-HSP27 (Ser82) Antibody #2401
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PhosphoSitePlus® protein, site, and accession data: HSP27
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IHC-P IHC-F IF-IC | H M R Mk | Endogenous | 27 | Rabbit |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
IHC-F=Immunohistochemistry (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
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 animals with a synthetic phospho- peptide corresponding to residues surrounding Ser82 of human HSP27. Antibodies are purified by protein A and peptide affinity chromatography.
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 42ÂșC for 0-2 hours as indicated.
Western Blotting
Western blot analysis of extracts of various cell lines, using Phospho-HSP27 (Ser82) Antibody #2401 (upper) or control HSP27 Antibody #2402 (lower).
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)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-HSP-27 (Ser82) Antibody in the presence of control peptide (left) or antigen specific peptide (right).
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft, showing cytoplasmic localization using Phospho-HSP27 (Ser82) Rabbit mAb.
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® #4084 (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 HSPs, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the HSP27 expression increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78, and Ser82 by MAPKAPK-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).
- Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
- Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
- Rouse, J. et al. (1994) Cell 78, 1027-1037.
- Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
- Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
- Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.
Application References
- Sarkar, D. et al. (2002) mda-7 (IL-24) mediates selective apoptosis in human melanoma cells by inducing the coordinated overexpression of the GADD family of genes by means of p38 MAPK. Proc. Natl. Acad. Sci. USA 99, 10054-10059. Applications: Western Blotting
- Hofmann, M. et al. (2004) Mechanical pressure-induced phosphorylation of p38 mitogen-activated protein kinase in epithelial cells via Src and protein kinase C. Biochemical and Biophysical Research Communications 316, 673-679. Applications: IC-IF Western Blotting
- Kennett, S. B. et al. (2004) Requirement of protein kinase C micro activation and calpain-mediated proteolysis for arachidonic acid-stimulated adhesion of MDA-MB-435 human mammary carcinoma cells to collagen type IV. J. Biol. Chem. 279, 3300-3307. Applications: Western Blotting
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Companion Products
- 2402 HSP27 (G31) Mouse mAb
- 2442 HSP27 Antibody (Rodent Preferred)
- 2405 Phospho-HSP27 (Ser78) Antibody
- 2404 Phospho-HSP27 (Ser15) Antibody
- 2406 Phospho-HSP27 (Ser82) Antibody II
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- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 8112 SignalStain® Antibody Diluent
For Research Use Only. Not For Use In Diagnostic Procedures.