Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to NFkB p105/ p50 aa 300-400.
WB: HeLa, MCF-7 and PC-12 cell lysates.
IHC-P: human prostate carcinoma and bladder carcinoma tissues.
This product is a recombinant rabbit monoclonal antibody.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.
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 [E381] images
Western blot - Anti-NFkB p105 / p50 antibody [E381] (ab32360)
Predicted band size : 50 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: NFkB p105 / p50 knockout HAP1 cell lysate (20 µg) Lane 3: HeLa cell lysate (20 µg) Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and 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 - Anti-NFkB p105 / p50 antibody [E381] (ab32360)
Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg
Secondary Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
References for Anti-NFkB p105 / p50 antibody [E381] (ab32360)
This product has been referenced in:
Liu Z et al. Mark4 promotes oxidative stress and inflammation via binding to PPAR? and activating NF-?B pathway in mice adipocytes. Sci Rep6:21382 (2016).
Read more (PubMed: 26888669) »
Vogel CF et al. Transgenic Overexpression of Aryl Hydrocarbon Receptor Repressor (AhRR) and AhR-Mediated Induction of CYP1A1, Cytokines, and Acute Toxicity. Environ Health Perspect124:1071-83 (2016).
Read more (PubMed: 26862745) »