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Chicken polyclonal antibody to Heat Shock Protein 27 (HSP27)


Catalogue No. C-1700-100
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.
Batch No. See vial label
Unit size 100 uL
Antigen Recombinant full length purified HSP27 from E. coli.
Antibody Type Polyclonal
Produced in Chicken
Applications Western Blotting (WB) and Immunocytochemistry (ICC). A dilution of 1:2,000 - 1:5,000 is recommended for WB. A dilution of 1:1,000 - 1:2,000 is recommended for ICC. 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, rat, mouse, porcine and bovine. It is expected that it will work on other mammal tissues.
Antibody Against Heat Shock Protein 27 (HSP27)
Form Lyophilised chicken IgY with 5% trehalose and 0.5% sodium azide.
Appearance White powder
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)
Chicken polyclonal antibody to Heat Shock Protein 27 (HSP27) Crude extract of HeLa cells. The antibody recognizes the ~27 kDa protein.
Chicken polyclonal antibody to Heat Shock Protein 27 (HSP27) HeLa cells stained with C-1700-100 (green) and a monoclonal antibody to High mobility Group B protein 1 (M-1702-100, red). The HSP27 antibody reveals strong cytoplasmic staining and penetrates into the actin rich ruffled margins. The blue stain reveals DNA in the nuclei of these cells.
Chicken polyclonal antibody to Heat Shock Protein 27 (HSP27) Left: Analysis of HSP27 expression in HeLa cells by Immunocytochemistry with chicken antibody to HSP27 (1:1,000, green). Cells were co-stained with a mouse antibody to HSP60 (red). Blue: DAPI nuclear stain. The HSP27 antibody produces strong cytoplasmic staining, while the HSP60 antibody specifically labels mitochondria. Right: Western blot analysis of whole cell lysates with chicken antibody to HSP27 (1:2,000, red, lanes 2-7). [1] protein standard, [2] NIH-3T3 , [3] C6, [4] HeLa, [5] HEK293, [6] SH-SY5Y, [7] COS-1. A strong, single band at 27 kDa corresponds to the HSP27 protein expressed in human cell lines. The antibody does not recognize rodent HSP27. The blot was simultaneously probed with a mouse antibody to beta-tubulin (green).
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