Revision 1

#8219

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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 Mol. Wt Isotype/Source
Phospho-IkappaB alpha (Ser32/36) (5A5) Mouse Monoclonal Antibody9246100 µl40 kDaMouse IgG1
IkappaB alpha (L35A5) Mouse Monoclonal Antibody (Amino-terminal Antigen)4814100 µl39 kDaMouse IgG1

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

UniProt ID:
#P25963

Entrez-Gene Id:
4792

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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).

Trademarks and Patents

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

PhosphoPlus 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

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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 1: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Western blot analysis of extracts from NIH/3T3 cells, untreated or TNF-α-treated (#8902, 20 ng/ml) for 5 minutes, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper) or IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814, and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 2: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper) and IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (lower).
undefined Image 3: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
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 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines, untreated or treated with IFNa (#36000, 20 ng/mL, 5 min); using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246 (upper), IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 4: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Immunoprecipitation of IkBa from HeLa cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is IκBα (L35A5) Mouse mAb (Amino-terminal Antigen), #4814. Western blot was performed using IκBα Antibody #9242.
undefined Image 5: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24 h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb (upper) and IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) #4814 (lower).
undefined Image 6: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
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 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human leiomyoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
undefined Image 7: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
undefined Image 8: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human renal adenocarcinoma, using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen).
undefined Image 9: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
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 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells, untreated (left), or TNF-α-treated (#8902, 10 ng/ml for 15 min, right) using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 10: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Flow cytometric analysis of NIH/3T3 cells, treated with TNF-α (#8902, 10 ng/ml for 5 min; blue, negative) or untreated (green, positive) using IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
undefined Image 11: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
Simple Western™ analysis of lysates (1 mg/mL) from serum-starved HeLa cells treated with human TNF⍺ (20ng/mL, 5min) using Phospho-IκBα (Ser32/36) (5A5) Mouse mAb #9246. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 12: PhosphoPlus<sup>®</sup> IkappaB alpha (Ser32/Ser36) Antibody Duet
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.