Western blot analysis of extracts from various cell lines, untreated or treated with 10uM MG132, using NIK Antibody #4994.
Western blot analysis of extacts from HeLa cells, untransfected or transfected with HA-NIK, using NIK Antibody #4994.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
NIK Antibody detects endogenous levels of total NIK protein.
Rat, Monkey, Bovine, Horse
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues adjacent to glycine 659 of human NIK. Antibodies are purified by protein A and peptide affinity chromatography.
Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, c-Rel, RelB, NF-κB1 (p105/p50), and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. Rel proteins bind p50 and p52 to form dimeric complexes that bind DNA and regulate transcription. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins (3-5). NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression (6-8). NIK and IKKα (IKK1) regulate the phosphorylation and processing of NF-κB2 (p100) to produce p52, which translocates to the nucleus (9-11).
Activation of NF-kappaB can be controlled by NF-kB-inducing kinase (NIK), a member of the MAP3K family that was originally identified as a TRAF2-interacting protein and thereby coupled to receptor activation (12). NIK forms a complex with and phosphorylates IKK1 and IKK2, subsequently leading to the phosphorylation of IkappaB and translocation of NF-kappaB to the nucleus (13-15).
Explore pathways + proteins related to this product.