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Mouse monoclonal antibody to Heat Shock Protein 27 (HSP27) [6H11]: IgG


Catalogue No. M-1692-100
Description The heat shock proteins were discovered, as the name suggests, since they are heavily upregulated when cells are stressed by temperatures above the normal physiological range. They are expressed in unstressed cells also and have a normal function as chaperones, helping other proteins to fold correctly, and are required in much greater amounts if the cell or tissue is stressed by heat. The increased levels are generated transcriptionally under the influence of a powerful transcription factor, the heat shock factor 1 (HSF1). The different heat shock proteins were originally named based on their SDS-PAGE mobility, so HSP27 has an apparent molecular weight of 27kDa. It is an abundant protein even under non-stress conditions and frequently shows up as a major spot on 2 dimensional gels of cells or tissues. It is known to associate with a variety of other proteins such as actin, intermediate filament subunits and ubiquitin and is found both in the cytoplasm and the nucleus of cells. HSP27 can become heavily phosphorylated under the influence of multiple protein kinases particularly as a result of activation of the p38/SAPK pathway. Upregulation of this protein is protective against neurodegenerative diseases at least in certain mouse models (1). Point mutations in the HSP27 gene are associated with two neurological diseases, Charcot-Marie-Tooth disease type 2F and distal hereditary motor neuropathy IIB (2). These diseases are associated with axonal loss apparently following defects in the transport of neurofilaments.
Unit size 100 ug
Antigen HSP27
Isotype IgG1
Clone 6H11
Produced in Mouse
Purity IgG
Applications Western Blotting (WB) and Immunocytochemistry (IC). A dilution of 1:5,000 - 1:10,000 is recommended for WB. A dilution of 1:500 - 1:1,000 is recommended for IC. Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity The antibody reacts with a 27 kDa band by Western blot on a crude extract from HeLa cells. It has also been used successfully for immunocytochemistry.
Species Against Human.
Antibody Against Heat Shock Protein 27 (HSP27)
Cross-reactivity Does not react with rodent protein.
Form Lyophilised with 5% trehalose
Reconstitution Reconstitute in sterile distilled water. Centrifuge to remove any insoluble material.
Storage After reconstitution of lyophilised antibody, aliquot and store at -20C for a higher stability. Avoid freeze-thaw cycles.
Expiry Date 12 months after purchase
General References 1. Wyttenbach, A et al. Heat shock protein 27 prevents cellular polyglutamine toxicity and suppresses the increase of reactive oxygen species caused by huntingtin. Hum. Molec. Genet. 11:1137-1151 (2002).
2. Evgrafov, OV et al. Mutant small heat-shock protein 27 causes axonal Charcot-Marie-Tooth disease and distal hereditary motor neuropathy. Nature Genet. 36:602-606 (2004).
Images (click to zoom)
Mouse monoclonal antibody to Heat Shock Protein 27 (HSP27) [6H11]: IgG Crude extract of HeLa cells. The antibody recognizes the 27 kDa protein.
Mouse monoclonal antibody to Heat Shock Protein 27 (HSP27) [6H11]: IgG HeLa cells stained with M-1692-100 (red) and also with our chicken polyclonal antibody to Vimentin, (C-1409-50). The HSP27 antibody reveals strong cytoplasmic staining and penetrates into the actin rich ruffled margins. The vimentin antibody stains a quite different cytoskeletal network, the intermediate filaments. The blue stain reveals DNA in the nuclei of these cells.
Mouse monoclonal antibody to Heat Shock Protein 27 (HSP27) [6H11]: IgG Left: Analysis of HSP27 expression (green) in HeLa cells by Immunocytochemistry. HSP27 was stained with mouse antibody at 1:2,000 dilution, and cells were co-stained with a chicken antibody to HSP60 (red). Blue: DAPI nuclear stain. The HSP27 antibody produces strong cytoplasm staining, while the HSP60 antibody specifically labels mitochondria. Right: Western blot analysis of HSP27 expression (green) in tissue or whole cell lysates using mouse anti-HSP27 antibody at 1:10,000 dilution. [1] protein standard, [2] rat brain, [3] mouse brain, [4] NIH-3T3, [5] HEK293, [6] HeLa. The strong single band at 27 kDa corresponds to the HSP27 protein, detected only in human cell lines, since this antibody is human specific. The blot was simultaneously probed with a rabbit antibody to HSP60 (red, lanes 2-6).
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