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NF-kB p65 Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# ER0815

NF-kB p65 Rabbit Polyclonal Antibody

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

  • Zebrafish

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

NF-kB p65 Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Synthetic peptide within N-terminal human RELA.

Species Reactivity

Human, Mouse, Rat, Zebrafish

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 60 kDa

Positive Control

Hela cell lysate, A549 cell lysate, PC12 cell lysate, Mouse embryonic stem cell lysate, NIH/3T3 cell lysate, zebrafish lysates, human lung cancer tissue, human lung tissue, human spleen tissue, mouse spleen tissue, rat spleen tissue, zebrafish, Hela.

Conjugation

unconjugated

RRID

AB_3069017

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:200

  • FC

  • 1:50-1:100

Target

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 and the heterodimeric p65-p50 complex appears to be most abundant one. In unstimulated cells, NF-κB is sequestered in the cytoplasm by IκB inhibitory proteins. NF-κB-activating agents can induce the phosphorylation of IκB proteins, targeting them for rapid degradation through the ubiquitin-proteasome pathway and releasing NF-κB to enter the nucleus where it regulates gene expression.

Background References

1. "Duration of nuclear NF-kappaB action regulated by reversible acetylation." Chen L.F., Fischle W., Verdin E., Greene W.C.Science 293:1653-1657(2001)

2. "A novel protein overexpressed in hepatoma accelerates export of NF-kappa B from the nucleus and inhibits p53-dependent apoptosis." Higashitsuji H., Higashitsuji H., Nagao T., Nonoguchi K., Fujii S., Itoh K., Fujita J. Cancer Cell 2:335-346(2002)

3. "Breast cancer metastasis suppressor 1 functions as a corepressor by enhancing histone deacetylase 1-mediated deacetylation of RelA/p65 and promoting apoptosis." Liu Y., Smith P.W., Jones D.R. Mol. Cell. Biol. 26:8683-8696(2006)

4. "SIRT2 regulates NF-kappaB dependent gene expression through deacetylation of p65 Lys310." Rothgiesser K.M., Erener S., Waibel S., Luscher B., Hottiger M.O. J. Cell Sci. 123:4251-4258(2010)

Post-translational Modification

Ubiquitinated by RNF182, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.; Monomethylated at Lys-310 by SETD6. 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.; 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.

Subcellular Location

Nucleus, cytoplasm.

UNIPROT

SwissProt: Q04206 Human

SwissProt: Q04207 Mouse

Entrez Gene: 309165 Rat

Synonyms

Avian reticuloendotheliosis viral (v rel) oncogene homolog A antibody

MGC131774 antibody

NF kappa B p65delta3 antibody

NFKB3 antibody

Nuclear Factor NF Kappa B p65 Subunit antibody

Nuclear factor NF-kappa-B p65 subunit antibody

Nuclear factor of kappa light polypeptide gene enhancer in B cells 3 antibody

Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 antibody

OTTHUMP00000233473 antibody

OTTHUMP00000233474 antibody

Expand

Images

  • Western blot analysis of NF-κB p65 on different lysates using anti-NF-κB p65 antibody at 1/1,000 dilution.<br /><span style="font-weight: bold;">Positive control:</span><br />Lane 1: Hela cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: PC12 cell lysate<br />Lane 4: Mouse embryonic stem cell lysate<br />Lane 5: NIH/3T3 cell lysate

    Western blot analysis of NF-κB p65 on different lysates using anti-NF-κB p65 antibody at 1/1,000 dilution.
    Positive control:
    Lane 1: Hela cell lysate
    Lane 2: A549 cell lysate
    Lane 3: PC12 cell lysate
    Lane 4: Mouse embryonic stem cell lysate
    Lane 5: NIH/3T3 cell lysate

  • Western blot analysis of NF-kB p65 on zebrafish lysates with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/2,000 dilution.<br /><br />Lysates at 20 µg/Lane.<br /><br />Predicted band size: 60kDa<br />Observed band size: 70 kDa<br /><br />Exposure time: 3min20s;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of NF-kB p65 on zebrafish lysates with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/2,000 dilution.

    Lysates at 20 µg/Lane.

    Predicted band size: 60kDa
    Observed band size: 70 kDa

    Exposure time: 3min20s;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ER0815) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

  • Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

  • Immunohistochemical analysis of paraffin-embedded zebrafish with Rabbit anti-NF-kB p65 antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ER0815" style="font-weight: bold;text-decoration: underline;">ER0815</a>) at 1/200 dilution for 1 hour at room temperature. 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.

    Immunohistochemical analysis of paraffin-embedded zebrafish with Rabbit anti-NF-kB p65 antibody (ER0815) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ER0815) at 1/200 dilution for 1 hour at room temperature. 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.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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