05-1469 | Anti-NFĸB Antibody, p65, clone 1G10.2

05-1469
200 µg  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, M, R, BWB, IF, ICCMPurifiedMonoclonal Antibody
      Description
      Catalogue Number05-1469
      DescriptionAnti-NFĸB Antibody, p65, clone 1G10.2
      Background InformationNFĸB (Nuclear Factor kappa B) represents a group of transcription factors that are structurally related and evolutionarily conserved proteins related to the proto-oncogene c-Rel. The five members in mammals includes Rel (cRel), RelA (p65), RelB, NFĸB1 (p50 and its precursor p105), and NFĸB2 (p52 and its precursor p100)1, 2. NFĸB/Rel proteins exist as homo- or heterodimer complexes.
      In a majority of unstimulated cells, the NFĸB transcription factors are bound to the inhibitory IĸB proteins and retained in the cytoplasm in their inactive form. Upon stimulation by multiple inducers, IĸBα is rapidly phosphorylated and degraded. This results in the release of the NFĸB complex, most commonly the p105/p65 heterodimer. The p105 subunit is cleaved into its active p50 form. This cleavage exposes the NLS sequence on the p50 subunit. The p50/p65 heteroduplex then translocates to the nucleus where it activates gene transcription.
      References
      Product Information
      FormatPurified
      PresentationPurified mouse monoclonal in 0.1M Tris-Glycine (pH 7.4) with 150mM NaCl and 0.05% NaN3.
      Applications
      ApplicationAnti-NFĸB Antibody, p65, clone 1G10.2 detects level of NFĸB & has been published & validated for use in WB, IF & IC.
      Key Applications
      • Western Blotting
      • Immunofluorescence
      • Immunocytochemistry
      Biological Information
      ImmunogenSynthetic peptide corresponding to the NLS region of human NFĸB p65.
      EpitopeNLS
      Clone1G10.2
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostMouse
      SpecificityDetects NFĸB p65.
      IsotypeIgG1κ
      Species Reactivity
      • Human
      • Mouse
      • Rat
      • Bovine
      Species Reactivity NoteHuman, Mouse, and Rat. Other species have not been tested. Expected to cross-react with bovine based on complete sequence homology.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Entrez Gene SummaryNFKB1 (MIM 164011) or NFKB2 (MIM 164012) is bound to REL (MIM 164910), RELA, or RELB (MIM 604758) to form the NFKB complex. The p50 (NFKB1)/p65 (RELA) heterodimer is the most abundant form of NFKB. The NFKB complex is inhibited by I-kappa-B proteins (NFKBIA, MIM 164008 or NFKBIB, MIM 604495), which inactivate NFKB by trapping it in the cytoplasm. Phosphorylation of serine residues on the I-kappa-B proteins by kinases (IKBKA, MIM 600664, or IKBKB, MIM 603258) marks them for destruction via the ubiquitination pathway, thereby allowing activation of the NFKB complex. Activated NFKB complex translocates into the nucleus and binds DNA at kappa-B-binding motifs such as 5-prime GGGRNNYYCC 3-prime or 5-prime HGGARNYYCC 3-prime (where H is A, C, or T; R is an A or G purine; and Y is a C or T pyrimidine).[supplied by OMIM]
      Purification MethodProtein G Purified
      UniProt Number
      UniProt SummaryFunction: NF-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. Ref.32

      Subunit structure: Component of the NF-kappa-B p65-p50 complex. Component of the NF-kappa-B p65-p50 complex. Homodimer; component of the NF-kappa-B p50-p50 complex. Component of the NF-kappa-B p105-p50 complex. Component of the NF-kappa-B p50-c-Rel complex. Component of a complex consisting of the NF-kappa-B p50-p50 homodimer and BCL3. Also interacts with MAP3K8. NF-kappa-B p50 subunit interacts with NCOA3 coactivator, which may coactivate NF-kappa-B dependent expression via its histone acetyltransferase activity. Interacts with DSIPI; this interaction prevents nuclear translocation and DNA-binding. Interacts with SPAG9 and UNC5CL. NFKB1/p105 interacts with CFLAR; the interaction inhibits p105 processing into p50. NFKB1/p105 forms a ternary complex with MAP3K8 and TNIP2. Interacts with GSK3B; the interaction prevents processing of p105 to p50. NFKB1/p50 interacts with NFKBIE. NFKB1/p50 interacts with NFKBIZ. Nuclear factor NF-kappa-B p50 subunit interacts with NFKBID By similarity.

      Subcellular location: Nucleus. Cytoplasm. Note= Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B).

      Induction: By phorbol ester and TNF-alpha.

      Domain: The 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 modification: While 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.

      Sequence similarities: Contains 7 ANK repeats.
      Contains 1 death domain.
      Contains 1 RHD (Rel-like) domain.
      Molecular Weight65 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely evaluated by Western Blot.

      Western Blot Analysis:
      This lot detected NFB, p65 at 1:1,000 dilution in Jurkat cell lysate resolved via SDS-PAGE and transferred to PVDF.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution.
      Packaging Information
      Material Size200 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-NFkB, p65, clone 1G10.2 - 2181277 2181277
      Anti-NFkB, p65, clone 1G10.2 - 2278555 2278555
      Anti-NFkB, p65, clone 1G10.2 - NG1726940 NG1726940
      Anti-NFkB, p65, clone 1G10.2 - NG1853752 NG1853752
      Anti-NFkB, p65, clone 1G10.2 -2851198 2851198
      Anti-NFkB, p65, clone 1G10.2 -2858456 2858456
      Anti-NFκB, p65, clone 1G10.2 - 2452691 2452691
      Anti-NFκB, p65, clone 1G10.2 -2680242 2680242
      Anti-NFκB, p65, clone 1G10.2 -2736494 2736494
      Anti-NFκB, p65, clone 1G10.2 Monoclonal Antibody 2982340

      Brochure

      Title
      Pathways and Biomarkers of Toll-like Receptor (TLR) Signaling

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      Categories

      Life Science Research > Antibodies and Assays > Primary Antibodies