Revision 6

#7946

Store at +4C

PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M

UniProt ID:
#Q16539, #O15264, #P53778, #Q15759

Entrez-Gene Id:
#1432, #5603, #6300, #5600

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
Phospho-p38 MAPK (T180/Y182) Mouse Ab Coated Microwells8125696 tests+4C
p38 MAPK Detection Rabbit mAb326131 eaGreen (Lyophilized)+4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)132721 eaRed (Lyophilized)+4C
Detection Antibody Diluent1333911 mlGreen+4C
HRP Diluent1351511 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
ELISA Sample Diluent1108325 mlBlue+4C
Cell Lysis Buffer (10X)980315 ml-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

Description

The PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-p38 MAP kinase phosphorylated at Thr180/Tyr182. A phospho-p38 MAP kinase (Thr180/Tyr182) mouse antibody has been coated onto the microwells. After incubation with cell lysates, phospho-p38 MAP kinase (Thr180/Tyr182) protein is captured by the coated antibody. Following extensive washing, a p38 MAP kinase rabbit detection antibody is added to detect the captured phospho-p38 MAP kinase (Thr180/Tyr182). Anti-rabbit IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-p38 MAP kinase (Thr180/Tyr182).

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit detects endogenous levels of p38 MAP kinase phosphorylated at Thr180/Tyr182 in human and mouse cells. As shown in Figure 1, a significant induction of p38 MAP kinase phosphorylation at Thr180/Tyr182 can be detected in HeLa cells using this Sandwich ELISA Kit following treatment with anisomycin or UV irradiation. The level of total p38 MAP kinase remains unchanged as shown by western analysis (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). Four isoforms of p38 MAPK, p38α, β, γ (also known as Erk6 or SAPK3), and δ (also known as SAPK4) have been identified. Similar to the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses, including osmotic shock, inflammatory cytokines, lipopolysaccharide (LPS), UV light, and growth factors (1-5). MKK3, MKK6, and SEK activate p38 MAPK by phosphorylation at Thr180 and Tyr182. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase 2 (3) and to phosphorylate the transcription factors ATF-2 (5), Max (6), and MEF2 (5-8). SB203580 (4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)-imidazole) is a selective inhibitor of p38 MAPK. This compound inhibits the activation of MAPKAPK-2 by p38 MAPK and subsequent phosphorylation of HSP27 (9). SB203580 inhibits p38 MAPK catalytic activity by binding to the ATP-binding pocket, but does not inhibit phosphorylation of p38 MAPK by upstream kinases (10).

Background References

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 6

#7946

PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit

Cell Signaling Technology Logo
Figure 1. Treatment of HeLa cells with anisomycin or UV irradiation stimulates phosphorylation of p38 MAP kinase at Thr180/Tyr182 as detected by the PathScan® Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit #7946, but does not affect the level of total p38 MAP kinase. HeLa cells were treated with 5 μg/ml anisomycin for 30 minutes at 37ºC or 45 mJ/cm2 UV irradiation followed by a 30 minute recovery period at 37ºC. The absorbance readings at 450 nm are shown in the top figure, while the western blots using p38 MAPK Antibody #9212 (left) or Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (right) are shown in the bottom figure.
ELISA Image 1: PathScan<sup>®</sup> Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit
Figure 2. The relationship between lysate protein concentration from untreated, anisomycin-treated or UV-irradiated HeLa cells and the absorbance at 450 nm is shown.
ELISA Image 2: PathScan<sup>®</sup> Phospho-p38 MAPK (Thr180/Tyr182) Sandwich ELISA Kit
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.