More info
Assay Range | 62.5--4000 pg/mL |
Sensitivity | 5.0 pg/mL |
Size | 96T |
Storage | Store at 2 - 8ºC. Keep reconstituted standard and detection Ab at -20 ºC |
Assay Principle | Sandwich ELISA |
Sample volume | 100 µL final volume, dilution factor varies on samples |
Detection Method | Chromogenic |
Kit Components
1. Recombinant Human HSP27 standard: 2 vials
2. One 96-well plate precoated with anti- Human HSP27 Ab
3. Sample diluent buffer: 12 mL - 1
4. Detection antibody: 130 µL, dilution 1:100
5. Streptavidin-HRP: 130 µL, dilution 1:100
6. Antibody diluent buffer: 12 mL x1
7. Streptavidin-HRP diluent buffer: 12 mL x1
8. TMB developing agent: 10 mL x1
9. Stop solution: 10 mL x1.
10. Washing solution (20x): 25 mL x1.
Background
Heat shock protein 27 (Hsp27), also known as heat shock protein beta-1 (HSPB1), is a protein encoded by the HSPB1 gene in humans. Hsp27 functions as a molecular chaperone belonging to the small heat shock protein (sHsp) family which includes other proteins such as ubiquitin, α-crystallin, Hsp20. Structurally, sHsp is characteristic of a homologous and highly conserved amino acid sequence, the so-called α-crystallin-domain at the C-terminus and a WD/EPF domain at N-terminus. HSP27 exists as a large oligomer. The WD/EPF domain is essential for the development of high molecular oligomers. The oligomerization depends on the physiology of the cells, the phosphorylation status of Hsp27 and the exposure to stress. Stress induces an increase of expression and phosphorylation of Hsp27. Stimulation of the p38 MAP kinase cascade by various agents, mitogens, inflammatory cytokines such as TNFα and IL-1β, hydrogen peroxide and other oxidants, leads to the activation of MAPKAP kinases 2 and 3 which directly phosphorylate mammalian sHsp. In response to an apoptotic stimulus, Hsp27 regulates apoptosis through direct interaction with cytochrome c to prevent the proper assembly of the apoptosome and subsequently the activation of procaspase9 and procaspase3.