The Total HSP27 Sandwich ELISA kit is a qualitative, solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total HSP27 protein. An HSP27 antibody has been coated onto the microwells. After incubation with cell lysates, both nonphospho- and phospho-HSP27 are captured by the coated antibody. Following extensive washing, an HSP27 mouse monoclonal antibody is added to detect the captured HSP27 protein. HRP-linked anti-mouse antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total HSP27 protein. Antibodies in this kit are custom formulations specific to the kit.
Heat shock protein (HSP) 27 is one of the small Hsps that are constitutively expressed at different levels in various cell types and tissues. Like other small heat shock proteins, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the expression level of HSP27 increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, 78 and 82 by MAPKAP kinase 2 as a result of the activation of the p38 MAP kinase pathway (2,3). Phosphorylation of HSP27 causes a change in its tertiary structure, which shifts from large homotypic multimers to dimers and monomers (4). It has been shown that phosphorylation and increased concentration of HSP27 modulates actin polymerization and reorganization (5,6)
This ELISA kit that detects endogenous levels of total HSP27 protein. Using Phospho-HSP27(Ser78) Sandwich ELISA Kit, a significant induction of phospho-HSP27 (Ser78) in HeLa cells treated with UV light can be detected. However, the level of total HSP27 (phospho and non-phospho), detected by this kit remains unchanged see (Figure 1). COS cells treated with UV light show similar results.
Human and monkey
H1830-55A1: Microtiter Plate, 1x96 wells (12 strips of 8 wells). Coated with HSP27 antibody
H1830-55A2: HSP27 Detection Antibody, 1x11ml. Green solution.
H1830-55A3: IgG (HRP), 1x11ml. Anti-mouse IgG linked to HRP. Red solution.
H1830-55A4: TMB Substrate, 1x11ml. Colorless solution.
H1830-55A5: Stop Solution, 1x11ml. Colorless solution
H1830-55A6: Wash Buffer, 20X, 1x25ml. Colorless solution.
H1830-55A7: Sample Diluent, 1x25ml. Blue solution.
H1830-55A8: Cell Lysis Buffer, 10X, 1x15ml. Yellowish solution.
Storage and Stability:
Store H1830-55A8 Cell Lysis Buffer, 10X at -20 degrees C Store other components at 4 degrees C. Kit is stable for up to 6 months.