Western blot analysis of extracts from differentiated THP-1 cells, untreated or LPS-treated for 15 minutes, using Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb (upper), IKKα Antibody #2682 (middle) and IKKβ (2C8) Rabbit mAb #2370 (lower).
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.
Western blot analysis of extracts from HeLa cells, treated with TNF-α for the indicated times, using Phospho-IKKα/β (Ser176/180) Antibody II (upper) or IKKβ Antibody #2684 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Immunohistochemical analysis of paraffin-embedded human gall bladder (chronic cholecystitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic localization, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung (chronic bronchitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.
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).
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.
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.
|Phospho-IKKα (Ser176)/IKKβ (Ser177) (C84E11) Rabbit mAb 2078||20 µl||
||H M||85 (IKKalpha), 87 (IKKbeta)||Rabbit|
|Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb 2697||20 µl||
||H Mk M R||85 IKK-alpha 87 IKK-beta||Rabbit IgG|
|Phospho-IKKα/β (Ser176/180) Antibody II 2694||20 µl||
||H Mk M R||85 IKK-alpha 87 IKK-beta||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Phospho-IKKalpha/beta (Ser176/180) Antibody Sampler Kit contains reagents to examine protein levels of IKKalpha when phosphorylated at Ser176/180 and IKKbeta when phosphorylated at Ser177/181. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.
Phospho-IKKα/β (Ser176/180) Antibody, Phospho-IKKα/β (Ser176/180) Antibody II, and Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb detect IKKα only when phosphorylated at Ser176/180 and IKKβ only when phosphorylated at Ser177/181.
Polyclonal antibodies #2687 and #2694 are produced by immunizing rabbits with a synthetic phosphopeptide corresponding to residues surrounding Ser176/180 of human IKKα and Ser177/181 of IKKβ, respectively, and are purified by protein A and peptide affinity chromatography. Monoclonal antibody #2697 is produced by immunizing rabbits with a synthetic phosphopeptide corresponding to residues surrounding Ser176/180 of human IKKα.
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).
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