Revision 2

#15401

Store at +4C

FastScan Total p38 alpha MAPK ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M R Mk

UniProt ID:
#Q16539

Entrez-Gene Id:
#1432

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes Product # Quantity Color
FastScan™ ELISA Microwell Strip Plate, 96 Well5325796 tests
p38α MAPK Rabbit Capture mAb318801 eaGreen (Lyophilized)
p38α MAPK Mouse HRP-linked mAb282271 eaRed (Lyophilized)
FastScan™ ELISA Capture Antibody Diluent160763 mlGreen
FastScan™ ELISA HRP Antibody Diluent281203 ml
TMB Substrate700411 ml
STOP Solution700211 ml
Sealing Tape545031 ea
ELISA Wash Buffer (20X)980125 ml
FastScan™ ELISA Cell Extraction Buffer (5X)6990510 ml
FastScan™ ELISA Cell Extraction Enhancer Solution (50X)252431 ml
FastScan™ ELISA Kit #15401 Positive Control Type 2334301 ea

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 FastScan™ Total p38α MAPK ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of p38α MAPK protein. To perform the assay, sample is incubated with a capture antibody conjugated with a proprietary tag and a second detection antibody linked to HRP, forming a sandwich with p38α MAPK in solution. This entire complex is immobilized to the plate via an anti-tag antibody. The wells are then washed to remove unbound material. TMB is then added. The magnitude of observed signal is proportional to the quantity of p38α MAPK protein.

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

IMPORTANT: This FastScan™ ELISA Kit requires 4 washes at Step 6 of the protocol.

Specificity/Sensitivity

The FastScan™ Total p38 alpha MAPK ELISA Kit detects endogenous levels of p38α MAPK, as shown in Figure 1. This kit does not cross-react with other p38 MAPK isoforms, such as β and γ, as shown in Figure 2. 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).

Trademarks and Patents

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

FastScan™ ELISA is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patents 9,086,407, 9,261,500, and 9,476,874, foreign equivalents, and child patents deriving therefrom.

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

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

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 2

Cell Signaling Technology Logo
Figure 1. The relationship between lysate protein concentration from HeLa cells and the absorbance at 450 nm using the FastScan™ Total p38α MAPK ELISA Kit #15401 is shown. Untreated HeLa cells were collected at 90% confluence and lysed.
ELISA Image 1: FastScan<sup>™</sup> Total p38 alpha MAPK ELISA Kit
Figure 2. The FastScan™ Total p38α MAPK ELISA Kit #15401 detects p38α MAPK protein, but not p38γ MAPK or p38β MAPK proteins. The absorbance at 450 nm for a panel of His or GST-tagged recombinant protein p38 MAPK isoforms (5 μg/mL for each) using the FastScan™ Total p38α MAPK ELISA Kit #15401 is shown in the upper figure. The corresponding western blots using a p38 MAPK antibody that recognizes p38γ, p38β, and p38α MAPK proteins (left panel) and a p38α MAPK antibody (right panel) are shown in the lower figure.
ELISA Image 2: FastScan<sup>™</sup> Total p38 alpha MAPK 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.