Product Pathways - PathScan ELISA
PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit #7290
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|Kit Includes||Volume||Solution Color|
|HSP27 Mouse mAb Coated Microwells||96 tests|
|P-HSP27 (S78) Rabbit Detection Antibody||11 ml|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
CST's PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-HSP27 (Ser78) protein. HSP27 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, HSP27 protein (phosphorylated and nonphosphorylated) is captured by the coated antibody. Following extensive washing, Phospho-HSP27 (Ser78) Antibody is added to detect the captured phospho-HSP27 (Ser78) protein. HRP-linked, anti-rabbit antibody #7074 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 phospho- HSP27 (Ser78) protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Phospho-HSP27 (Ser78) Sandwich ELISA Kit detects endogenous levels of phospho-HSP27 (Ser78) protein. Using this Sandwich ELISA Kit #7290, a significant induction of phospho-HSP27 (Ser78) can be detected in HeLa cells treated with UV light. However, the level of total HSP27 (phospho and non-phospho), detected by the Total HSP27 Sandwich ELISA Kit #7295, remains unchanged (Figure 1). COS cells treated with UV light show similar results (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of HeLa cells with UV light stimulates phosphorylation of HSP27 at Ser78 detected by PathScanTM Phospho-HSP27 (Ser78) Sandwich ELISA kit, #7290, but does not affect the level of total HSP27 protein detected by PathScanTM Total HSP27 Sandwich ELISA kit, #7295. The corresponding Western blots, using Phospho-HSP27 (Ser78) Antibody #2405 (right) or HSP27 (G31) Monoclonal Antibody #2402 (left), are also shown.
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 HSPs, HSP27 is regulated at both the transcriptional and posttranslational levels (1). In response to stress, the HSP27 expression increases several-fold to confer cellular resistance to the adverse environmental change. HSP27 is phosphorylated at Ser15, Ser78, and Ser82 by MAPKAPK-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).
- Arrigo, A.P. and Landry, J. (1994) Cold Spring Harbor Laboratory Press, NY, 335-373.
- Landry, J. et al. (1992) J. Biol. Chem. 267, 794-803.
- Rouse, J. et al. (1994) Cell 78, 1027-1037.
- Rogalla, T. et al. (1999) J. Biol. Chem. 274, 18947-18956.
- Lavoie, J. et al. (1993) J. Biol. Chem. 268, 24210-24214.
- Rousseau, S. et al. (1997) Oncogene 15, 2169-2177.
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- 7295 PathScan® Total HSP27 Sandwich ELISA Kit
- 7152 PathScan® Phospho-HSP27 (Ser82) Sandwich ELISA Kit
- 2405 Phospho-HSP27 (Ser78) Antibody
- 9709 Phospho-HSP27 (Ser82) (D1H2) XP® Rabbit mAb
- 2402 HSP27 (G31) Mouse mAb
- 2442 HSP27 Antibody (Rodent Preferred)
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.