Revision 4

#72950

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FastScan Total IκBα ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M R Mk

UniProt ID:
#P25963

Entrez-Gene Id:
#4792

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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
IκBα Rabbit Capture mAb233731 eaGreen (Lyophilized)
IκBα Mouse HRP-linked mAb589301 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 #72950 Positive Control Type 2868371 ea

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The FastScan™ Total IκBα ELISA Kit is a sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of IκBα. 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 IκBα 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 IκBα.

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

Specificity/Sensitivity

The FastScan™ Total IκBα ELISA Kit detects endogenous levels of IκBα, as shown in 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

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins (1-3). Activation occurs via phosphorylation of IκBα at Ser32 and Ser36 followed by proteasome-mediated degradation that results in the release and nuclear translocation of active NF-κB (3-7). IκBα phosphorylation and resulting Rel-dependent transcription are activated by a highly diverse group of extracellular signals including inflammatory cytokines, growth factors, and chemokines. Kinases that phosphorylate IκB at these activating sites have been identified (8).

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected

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 4

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Figure 1. Treatment of HeLa cells with TNF-α stimulates phosphorylation of IκBα at Ser32/36 whereas the level of total IκBα is decreased due to degradation. The relationship between lysate protein concentration from untreated and TNF-α -treated HeLa cells and the absorbance at 450 nm using the FastScan™ Total IκBα ELISA Kit #72950 is shown in the upper figure. The corresponding western blots using phospho-IκBα (Ser32/36) antibody (left panel) and IκBα antibody (right panel) are shown in the lower figure. After serum starvation, HeLa cells were treated with Human Tumor Necrosis Factor-α (hTNF-α) #8902 (10 ng/ml) for 5 minutes at 37°C, and then lysed.

ELISA Image 1: FastScan<sup>™</sup> Total IκBα 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.