NF-κB p65 Antibody (YA4176)

(Synonyms: NFkappaB p65; p65; NFKB3; RELA)
Customer Review

Based on 1 Customer Validation

NF-κB p65 Antibody (YA4176) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to NF-κB p65.

For research use only. We do not sell to patients.
  • Host:

    Mouse

  • Isotype:

    IgG

  • Application:

    WB, IHC-P, ELISA

  • Reactivity :

    Human, Mouse, Rat, Rabbit, Monkey

  • Formulation:

    Supplied in PBS with 0.05% sodium azide.

  • Conjugation:
    Non-conjugated

Applications

Application
WB Info
WB: Western Blot
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
ELISA Info
ELISA: Enzyme Linked Immunosorbent Assay
Dilution Ratio 1:500-1:2000 1:100-1:500 1:10000

Product Details

Description

NF-κB p65 Antibody (YA4176) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to NF-κB p65.

  • Host Mouse
  • Clonality Monoclonal
  • Species Reactivity
    Human, Mouse, Rat, Rabbit, Monkey
  • Observed Molecular Weight
    Observed band size: 65 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 60 kDa
Immunogen

Purified recombinant fragment of human NF-κB p65 aa 136-224.

Purification

affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG

RRID

AB_3718913

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in PBS with 0.05% sodium azide.

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Western blot analysis of extracts from A549(lane 2(20μg) , MCF-7 (lane 3(20μg) ,RAW264.7(lane 4(20μg)and Hela( lane 5(20μg) using NF-κB p65 Antibody(HY-P84479).Proteins were transferred to a PVDF membrane and blocked with 5% nonfat powdered milk in TBST for 2 hour at room temperature. The primary antibody ( 1/750) and Loading control antibody (Beta Actin, HY-P83730,1/10000) was used in 5% nonfat powdered milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001,1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human cervix tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human endometrium tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human adrenal gland tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human gallbladder tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Experimental Validation Results for NF-κB p65 Antibody (YA4176)
    Immunohistochemical analysis of paraffin-embedded human small intestine tissue using NF-κB p65 Antibody (HY-P84479, 1/300) . The section was pretreated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked with quick block buffer for 0.5 hours at room temperature, washed with PBS and PBST, and then incubated with the primary antibody overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Background

  • Function

    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. The heterodimeric RELA-NFKB1 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. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. 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. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Besides its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells. The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection

  • Subcellular Localization

    Nucleus; Cytoplasm

  • Isoforms & Post-Translational Modification

    NF-κB has 4 isomers: Q04206-1: 60219 Da (predicted); Q04206-2: 58807 Da (predicted); Q04206-3: 59068 Da (predicted); Q04206-4: 59910 Da (predicted).
    Ubiquitinated by RNF182, leading to its proteasomal degradation (PubMed:31432514). Degradation is required for termination of NF-kappa-B response. Polyubiquitinated via 'Lys-29'-linked ubiquitin; leading to lysosomal degradation (PubMed:21518757);Monomethylated at Lys-310 by SETD6 (PubMed:21515635). Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes (By similarity);Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity. Phosphorylation at Ser-276 by RPS6KA4 and RPS6KA5 promotes its transactivation and transcriptional activities;Phosphorylation at Ser-75 by herpes simplex virus 1/HHV-1 inhibits NF-kappa-B activation;Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3 and SIRT2. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation at Lys-310 promotes interaction with BRD4. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of Lys-310. Lys-310 is deacetylated by SIRT2;S-nitrosylation of Cys-38 inactivates the enzyme activity;Sulfhydration at Cys-38 mediates the anti-apoptotic activity by promoting the interaction with RPS3 and activating the transcription factor activity;Sumoylation by PIAS3 negatively regulates DNA-bound activated NF-kappa-B;Proteolytically cleaved within a conserved N-terminus region required for base-specific contact with DNA in a CPEN1-mediated manner, and hence inhibits NF-kappa-B transcriptional activity (PubMed:18212740)

  • Subunit

    Component of the NF-κB p65-p50 complex. Interacts with NFKBIA. Interacts with NFKBIE. Interacts with NFKBIZ. Interacts with EHMT1 (via ANK repeats). Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, ELP1 and MAP3K14. Interacts with HDAC3; HDAC3 mediates the deacetylation of RELA. Interacts with HDAC1; the interaction requires non-phosphorylated RELA. Interacts with CBP; the interaction requires phosphorylated RELA. Interacts (phosphorylated at 'Thr-254') with PIN1; the interaction inhibits p65 binding to NFKBIA. Interacts with SOCS1. Interacts with UXT. Interacts with MTDH and PHF11. Interacts with ARRB2. Interacts with NFKBIA (when phosphorylated), the interaction is direct; phosphorylated NFKBIA is part of a SCF(BTRC)-like complex lacking CUL1. Interacts with RNF25. Interacts (via C-terminus) with DDX1. Interacts with UFL1 and COMMD1. Interacts with BRMS1; this promotes deacetylation of 'Lys-310'. Directly interacts with MEN1; this interaction represses NFKB-mediated transactivation. Interacts with AKIP1, which promotes the phosphorylation and nuclear retention of RELA. Interacts (via the RHD) with GFI1; the interaction, after bacterial lipopolysaccharide (LPS) stimulation, inhibits the transcriptional activity by interfering with the DNA-binding activity to target gene promoter DNA. Interacts (when acetylated at Lys-310) with BRD4; leading to activation of the NF-κB pathway. Interacts with MEFV. Interacts (via N-terminus) with CPEN1. Interacts with FOXP3. Interacts with CDK5RAP3. Interacts with DHX9; this interaction is direct and activates NF-κB-mediated transcription. Interacts with LRRC25. Interacts with ZBTB7A; involved in the control by RELA of the accessibility of target gene promoters. Directly interacts with DDX3X; this interaction may trap RELA in the cytoplasm, impairing nuclear relocalization upon TNF activating signals.

  • SwissProt ID

    Q04206

  • Gene ID
  • Synonyms

    NFkappaB p65; p65; NFKB3; RELA

NF-κB p65 Antibody (YA4176) Related Classifications

MOQ
Minimum order quantity
100 mg

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