Preparation And Storage
Recommended Assay Procedures
Applications include western blot analysis and immunoprecipitation. In immunoprecipitation experiments, it is reported that the C73-764 antibody brings down the IKK complex (IKKα, IKKβ, and IKKγ). The immunoprecipitation application has not been evaluated at BD Pharmingen. NIH/3T3 cells (ATCC CRL 1658) are recommended as a western blot positive control.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
The transcription factor NF-κB, (nuclear factor kappa-B) is controlled by interaction with an inhibitory subunit, IκB, which restricts NF-κB to the cytoplasm. Following stimulation by various cytokines or other stimuli, IκB becomes degraded and NF-κB is released to the nucleus. The release of IκB from NF-κB is a critical step in the activation of NF-κB signal pathways. A group of proteins form an NF-κB regulatory complex, or signalsome. Two members of this complex are a pair of closely related serine/threonine kinases, IKKα and IKKβ (aka IKK-1 and IKK-2), which phosphorylate critical residues of IκB, thus targeting it for subsequent degradation. The IKK complex contains similar amounts of IKKα, IKKβ, as well as two other polypeptides, which are differentially processed forms of a third subunit, IKKγ. IKKα and IKKβ become activated following phosphorylation by upstream kinases, including NF-κB-inducing kinase (NIK) and MEKK1. IKKγ interacts preferentially with IKKβ and is required for the activation of the IKK complex. Thus the IKK kinases play an important role in the activation of NF-κB. IKKγ migrates as a doublet between 50-55 kDa in SDS-PAGE.
Development References (6)
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DiDonato JA, Hayakawa M, Rothwarf DM, Zandi E, Karin M. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB. Nature. 1997; 388(6642):548-554. (Biology). View Reference
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Li ZW, Chu W, Hu Y, et al. The IKKbeta subunit of IkappaB kinase (IKK) is essential for nuclear factor kappaB activation and prevention of apoptosis. J Exp Med. 1999; 189(11):1839-1845. (Clone-specific: Immunoprecipitation). View Reference
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Ling L, Cao Z, Goeddel DV. NF-kappaB-inducing kinase activates IKK-alpha by phosphorylation of Ser-176. Proc Natl Acad Sci U S A. 1998; 95(7):3792-3797. (Biology). View Reference
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Mercurio F, Zhu H, Murray BW, et al. IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation. Science. 1997; 278(5339):860-866. (Biology). View Reference
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Nakano H, Shindo M, Sakon S, et al. Differential regulation of IkappaB kinase alpha and beta by two upstream kinases, NF-kappaB-inducing kinase and mitogen-activated protein kinase/ERK kinase kinase-1. Proc Natl Acad Sci U S A. 1998; 95(7):3537-3542. (Biology). View Reference
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Rothwarf DM, Zandi E, Natoli G, Karin M. IKK-gamma is an essential regulatory subunit of the IkappaB kinase complex. Nature. 1998; 395(6699):297-300. (Biology). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
