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NF-kB p100 / NFKB2 Mouse Monoclonal Antibody [16B2]

Usd: 350

Specification

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Catalog# EM1901-78

NF-kB p100 / NFKB2 Mouse Monoclonal Antibody [16B2]

Application

  • WB

  • IHC-P

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

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

Overview

Product Name

NF-kB p100 / NFKB2 Mouse Monoclonal Antibody [16B2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human NFKB2 aa 1-50 / 900.

Product Specificity

NF-κB2 p100 (16B2) Mouse mAb detects endogenous levels of both the p100 precursor and the p52 active form of NFKB2.

Species Reactivity

Human

Validated Applications

WB, IHC-P, FC

Molecular Weight

Predicted band size: 97 kDa

Positive Control

Human tonsil tissue, human placenta tissue, A431 cell, Siha cell lysates.

Conjugation

unconjugated

Clone Number

16B2

RRID

AB_3068923

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, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:50-1:200

  • 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. 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 complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor.

Background References

1. Dobrzanski P. et. al. Differential interactions of Rel-NF-kappa B complexes with I kappa B alpha determine pools of constitutive and inducible NF-kappa B activity. EMBO J. 13:4608-4616(1994)

Post-translational Modification

While translation occurs, the particular unfolded structure after the GRR repeat promotes the generation of p52 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.; Subsequent to MAP3K14-dependent serine phosphorylation, p100 polyubiquitination occurs then triggering its proteasome-dependent processing.; Constitutive processing is tightly suppressed by its C-terminal processing inhibitory domain, named PID, which contains the death domain.

Subcellular Location

Nucleus, Cytoplasm.

UNIPROT

SwissProt: Q00653 Human

Synonyms

CVID10 antibody

DNA binding factor KBF2 antibody

H2TF1 antibody

Lymphocyte translocation chromosome 10 protein antibody

LYT 10 antibody

NF kB2 antibody

NFKB p52/p100 subunit antibody

Nuclear factor Kappa B subunit 2 antibody

Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 (p49/p100) antibody

Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of NF-kB p100 / NFKB2 on different lysates with Mouse anti-NF-kB p100 / NFKB2 antibody (<a href="/products/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-NF-kB p100 / NFKB2 KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 97 kDa<br />Observed band size: 120 kDa<br /><br />Exposure time: 60 seconds; ECL: K1801;<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/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of NF-kB p100 / NFKB2 on different lysates with Mouse anti-NF-kB p100 / NFKB2 antibody (EM1901-78) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-NF-kB p100 / NFKB2 KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 97 kDa
    Observed band size: 120 kDa

    Exposure time: 60 seconds; ECL: K1801;

    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 (EM1901-78) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of NF-kB p100 / NFKB2 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:5,000 dilution was used for 1 hour at room temperature.<br />Predicted band size: 97 kDa<br />Observed band size: 52/100 kDa

    Western blot analysis of NF-kB p100 / NFKB2 on Siha cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-78, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
    Predicted band size: 97 kDa
    Observed band size: 52/100 kDa

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-NF-kB p100 / NFKB2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>, 1/100) for 30 minutes 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 tonsil tissue using anti-NF-kB p100 / NFKB2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-78, 1/100) for 30 minutes 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 placenta tissue using anti-NF-kB p100 / NFKB2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>, 1/200) for 30 minutes 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 placenta tissue using anti-NF-kB p100 / NFKB2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-78, 1/200) for 30 minutes 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.

  • Flow cytometric analysis of NF-kB p100 / NFKB2 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/EM1901-78" style="font-weight: bold;text-decoration: underline;">EM1901-78</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488 Goat anti-Mouse IgG antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of NF-kB p100 / NFKB2 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-78, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488 Goat anti-Mouse IgG antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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

Citation

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metafield image

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