Revision 4

#36214

Store at -20C

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.

Applications:
W, IHC-P, FC-FP, ELISA

Reactivity:
H M R Hm Mk

Sensitivity:
Endogenous

MW (kDa):
85 IKK-alpha 87 IKK-beta

Source/Isotype:
Rabbit IgG

UniProt ID:
#O15111

Entrez-Gene Id:
1147

Product Usage Information

This product is the carrier free version of product #2697. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

Formulation

Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

For standard formulation of this product see product #2697

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity/Sensitivity

Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free) detects IKKα only when phosphorylated at Ser176/180 and IKKβ only when phosphorylated at Ser177/181.

Species predicted to react based on 100% sequence homology

Bovine

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser176/180 of human IKKα.

Background

The NF-κB/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins (1-3). Most agents that activate NF-κB do so through a common pathway based on phosphorylation-induced, proteasome-mediated degradation of IκB (3-7). The key regulatory step in this pathway involves activation of a high molecular weight IκB kinase (IKK) complex whose catalysis is generally carried out by three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase and IKKγ serves as the regulatory subunit (8,9). Activation of IKK depends upon phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (Ser176 and Ser180 in IKKα), which causes conformational changes, resulting in kinase activation (10-13).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

W: Western Blotting IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized) ELISA: ELISA

Cross-Reactivity Key

H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a 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 4

Cell Signaling Technology Logo
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-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Western Blotting Image 1: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Western blot analysis of extracts from HeLa cells, untreated or treated with TNFa (#16789, 20 ng/mL, 10 min) and Calyculin A (#9902, 100 nM, 10 min); THP-1 cells differentiated with TPA (#4174, 80 nM for 16hr) untreated and treated with LPS (#14011, 1 μg/ml for 1hr) using #2697 (upper) or IKKα (D3W6N) Rabbit mAb #61294, IKKβ (D30C6) Rabbit mAb #8943 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower). Data were generated using the standard formulation of this product.
Western Blotting Image 2: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Western blot analysis of extracts from TNF-alpha and Calyculin A treated HeLa and NIH/3T3 cells, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Western Blotting Image 3: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
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

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human gall bladder (chronic cholecystitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic localization, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
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

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb in the presence of control peptide (left) or Phospho-IKK-alpha/beta (Ser176/180) Blocking Peptide #1023 (right). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human lung (chronic bronchitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 5: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
Flow cytometric analysis of THP-1 cells, untreated (blue) and with TPA and LPS (green) using IKK-α (Ser176/Ser180) phosphate Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody. Data were generated using the standard formulation of this product.
Flow Cytometry Image 1: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
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

Cell Signaling Technology Logo
Data using Phospho-IKKα (Ser176/180) Matched Antibody Pair #67526 are shown. The two antibodies function as a capture-target-detection sandwich (#36214 and #49706, respectively) to detect endogenous protein expression in positive and negative controls.
ELISA Image 1: Phospho-IKK alpha/beta (Ser176/180) (16A6) Rabbit Monoclonal Antibody (BSA and Azide Free)
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.