Product NameHeat shock protein 27 (HSP-27), Mouse Monoclonal Antibody
Product DescriptiongoogleMouse anti-Heat shock protein 27 (HSP-27) Monoclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Antibody HostMouse
Antibody TypeMonoclonal
SpecificityThe 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. Does not react with rodent protein.
Species ReactivityHuman
Immunogen DescriptionRecombinant full length human HSP27 expressed in and purified from E. coli
Product DescriptionMouse anti-Heat shock protein 27 (HSP-27) Monoclonal Antibody (Unconjugated), suitable for WB, ICC.
Application(s)ICC, WB
Application DetailsWestern Blotting (WB) and Immunocytochemistry (ICC). 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.
TargetHeat shock protein 27 (HSP-27)
SpecificityThe 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. Does not react with rodent protein.
Target Host SpeciesHuman
Species ReactivityHuman
Antibody HostMouse
Antibody TypeMonoclonal
Antibody IsotypeIgG1
Clone Name6H11
ConjugateUnconjugated
Immunogen DescriptionRecombinant full length human HSP27 expressed in and purified from E. coli
Purity DescriptionIgG
FormatLyophilized from PBS buffer pH 7.2-7.6 with 0.1% trehalose, without preservatives
Reconstitution InstructionsSpin vial briefly before opening. Reconstitute in 100 uL sterile-filtered, ultrapure water. Centrifuge to remove any insoluble material.
Storage InstructionsAfter reconstitution of lyophilized antibody, aliquot and store at -20°C for a higher stability. Avoid freeze-thaw cycles.
Batch NumberPlease see item label.
Expiration Date12 months after date of receipt (unopened vial).
Scientific BackgroundThe 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 27 kDa. 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.
Crude extract of HeLa cells. The antibody recognizes the 27 kDa protein.
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.
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).
General ReferencesWyttenbach, 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). 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).
1800 605-5127
+61 (0)8 8352 7711
