The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Detects a band of approximately 105 kDa (predicted molecular weight: 105 kDa).
Use at an assay dependent dilution.
FunctionNF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and RelB-p50 complexes are transcriptional activators. The NF-kappa-B p50-p50 homodimer is a transcriptional repressor, but can act as a transcriptional activator when associated with BCL3. NFKB1 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p105 and generation of p50 by a cotranslational processing. The proteasome-mediated process ensures the production of both p50 and p105 and preserves their independent function, although processing of NFKB1/p105 also appears to occur post-translationally. p50 binds to the kappa-B consensus sequence 5'-GGRNNYYCC-3', located in the enhancer region of genes involved in immune response and acute phase reactions. In a complex with MAP3K8, NFKB1/p105 represses MAP3K8-induced MAPK signaling; active MAP3K8 is released by proteasome-dependent degradation of NFKB1/p105.
DomainThe C-terminus of p105 might be involved in cytoplasmic retention, inhibition of DNA-binding, and transcription activation. Glycine-rich region (GRR) appears to be a critical element in the generation of p50.
Post-translational modificationsWhile translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p50 making it an acceptable substrate for the proteasome. This process is known as cotranslational processing. The processed form is active and the unprocessed form acts as an inhibitor (I kappa B-like), being able to form cytosolic complexes with NF-kappa B, trapping it in the cytoplasm. Complete folding of the region downstream of the GRR repeat precludes processing. Phosphorylation at 'Ser-903' and 'Ser-907' primes p105 for proteolytic processing in response to TNF-alpha stimulation. Phosphorylation at 'Ser-927' and 'Ser-932' are required for BTRC/BTRCP-mediated proteolysis. Polyubiquitination seems to allow p105 processing. S-nitrosylation of Cys-61 affects DNA binding.
Cellular localizationNucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor.
Nuclear factor kappa B DNA binding subunit antibody
Nuclear factor kappa-B, subunit 1 antibody
Nuclear factor NF kappa B p105 subunit antibody
Nuclear factor NF kappa B p50 subunit antibody
Nuclear factor NF-kappa-B p50 subunit antibody
Nuclear factor of kappa light chain gene enhancer in B cells 1 antibody
Nuclear factor of kappa light polypeptide gene enhancer in B cells 1 antibody
Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 antibody
p84/NF-kappa-B1 p98 antibody
Transcription factor NFKB1 antibody
Anti-NFkB p105 / p50 antibody [13A2F] images
Western blot - Anti-NFkB p105 / p50 antibody [13A2F] (ab72138)
Predicted band size : 105 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: NFκB p105/p50 knockout HAP1 cell lysate (20 µg) Lane 3: CellX1 cell lysate (20 µg) Lane 4: CellX2 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab72138 observed at 50 and 120 kDa. Red - loading control, ab181602, observed at 37 kDa. ab72138 was shown to recognize NFκB p105/p50 when NFκB p105/p50 knockout samples were used, along with additional cross-reactive bands. Wild-type and NFκB p105/p50 knockout samples were subjected to SDS-PAGE. ab72138 and ab181602 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - NFkB p105 / p50 antibody [13A2F] (ab72138)
All lanes : Anti-NFkB p105 / p50 antibody [13A2F] (ab72138) at 1/2000 dilution
Lane 1 : HeLa cell lyaste Lane 2 : A431 cell lysate Lane 3 : K562 cell lysate Lane 4 : HepG2 cell lysate