Rabbit Multiclonal NFkB p105 / p50 phospho S337 antibody. Suitable for WB, ChIP and reacts with Human samples. Immunogen corresponding to Synthetic Peptide within Human NFKB1 phospho S337.
IgG
Rabbit
Preservative: 0.09% Sodium azide
Constituents: 99.91% PBS
Liquid
Multiclonal
WB | ChIP | |
---|---|---|
Human | Tested | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1-2 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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.Nuclear factor NF-kappa-B p105 subunitP105 is the precursor of the active p50 subunit (Nuclear factor NF-kappa-B p50 subunit) of the nuclear factor NF-kappa-B (PubMed:1423592). Acts as a cytoplasmic retention of attached NF-kappa-B proteins by p105 (PubMed:1423592).Nuclear factor NF-kappa-B p50 subunitConstitutes the active form, which associates with RELA/p65 to form the NF-kappa-B p65-p50 complex to form a transcription factor (PubMed:1740106, PubMed:7830764). Together with RELA/p65, 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 (PubMed:1740106, PubMed:7830764).
Nuclear factor NF-kappa-B p105 subunit, DNA-binding factor KBF1, EBP-1, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1, NFKB1
Rabbit Multiclonal NFkB p105 / p50 phospho S337 antibody. Suitable for WB, ChIP and reacts with Human samples. Immunogen corresponding to Synthetic Peptide within Human NFKB1 phospho S337.
IgG
Rabbit
Preservative: 0.09% Sodium azide
Constituents: 99.91% PBS
Liquid
Multiclonal
RP23040087
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains.
Recombinant multiclonal antibodies offer the sensitivity of polyclonal antibodies by recognising multiple epitopes, along with consistency of a recombinant antibody.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Enrichment of endogenous NFkB p50 pSer337 protein at specific gene loci using ab308112: Chromatin Immunoprecipitation (ChIP) was performed using ab308112, 5 μg on sheared chromatin from 2 million HeLa cells treated with 50 ng/mL of TNFalpha for 45 minutes. Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed with optimized PCR primer pairs for the promoter of active IL-8, IkB gene, used as positive control target, and the SAT2, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
Western blot analysis of NFkB p105 / p50 (phospho S337) in whole cell extracts from TNFa treated Jurkat (20 ng/mL, 15 min) using ab308112 at a dilution of 1 µg/mL. Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody followed by chemiluminescence (ECL). Results show a band at ~50 kDa.
All lanes: Western blot - Anti-NFkB p105 / p50 (phospho S337) antibody [RP23040087] - ChIP Grade (ab308112) at 1 µg/mL
All lanes: whole cell extracts from TNFa treated Jurkat cells
All lanes: HRP-conjugated Goat anti-Rabbit secondary antibody
Developed using the ECL technique.
Observed band size: 50 kDa
Western blot analysis was performed on whole cell extracts (30 µg lysate) of Jurkat (Lane 1), Jurkat treated with TNF alpha (Lane 2). The blots were probed with ab308112, 1-2 µg/mL and detected by chemiluminescence using a Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate (0.4 µg/mL, 1:2500 dilution). A 55 kDa band corresponding to NFkB p105 / p50 (phospho S337) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using a 4-12% Bis-Tris gel, electrophoresis system and pre-stained protein standard. Resolved proteins were then transferred onto a nitrocellulose membrane. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed (ECL).
All lanes: Western blot - Anti-NFkB p105 / p50 (phospho S337) antibody [RP23040087] - ChIP Grade (ab308112) at 1 µg/mL
Lane 1: Jurkat whole cell extract at 30 µg
Lane 2: Jurkat treated with TNF alpha at 30 µg
All lanes: Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP conjugate at 1/2500 dilution
Developed using the ECL technique.
Predicted band size: 105 kDa
Observed band size: 55 kDa
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