Overview

Introduction
Heat shock protein 27 (Hsp27) also known as heat shock protein beta-1 (HSPB1), is a protein that in humans is encoded by the HSPB1 gene. Hsp27 appears in many cell types, especially all types of muscle cells. It is located mainly in the cytosol, but also in the perinuclear region, endoplasmatic reticulum, and nucleus. It is overexpressed during different stages of cell differentiation and development. The main function of Hsp27 is to provide thermotolerance in vivo, cytoprotection, and support of cell survival under stress conditions. Another function of Hsp27 is the activation of the proteasome. It is also involved in the apoptotic signalling pathway.

Principle of the Assay  
This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti- HSP27 polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti- HSP27 polyclonal antibody was used as detection antibodies. The standards, test samples and biotin conjugated detection antibody were added to the wells subsequently, and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the HSP27 amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of HSP27 can be calculated.

Range: 62.5 pg/ml-4000 pg/ml

Sensitivity < 5 pg/ml

Storage and Expiration: Store at 2-8℃ for 4 months, or at -20℃ for 8 months.

Application: For quantitative detection of HSP27 in human serum, body fluids, tissue lysates or cell culture supernates. 

Kit components
1. One 96-well plate pre-coated with anti-Human HSP27 antibody
2. Lyophilized HSP27 standards: 2 tubes (10ng / tube)                     
3. Sample / Standard diluent buffer: 30ml
4. Biotin conjugated anti-Human HSP27 antibody (Concentrated): 130μl. Dilution: 1:100
5. Antibody diluent buffer: 12ml
6. Avidin-Biotin-Peroxidase Complex (ABC) (Concentrated): 130μl. Dilution: 1:100
7. ABC diluent buffer: 12ml
8. TMB substrate: 10ml
9. Stop solution: 10ml
10. Wash buffer (25X): 30ml

Note: Reconstitute standards and test samples with Kit Component 3.

Gene Information

OMIM: 602195

Swiss Port: P04792
Gene ID: 3315

Other Information

Please check datasheet for protocol.