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Cell Biology Apoptosis Intracellular NFkB p52
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

  • Datasheet
  • SDS
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Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
  • Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4686-66] to NFkB p100/NFKB2
  • Suitable for: ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-NFkB p100/NFKB2 antibody [EPR4686-66]
    See all NFkB p100/NFKB2 primary antibodies
  • Description

    Rabbit monoclonal [EPR4686-66] to NFkB p100/NFKB2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, WBmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Recombinant fragment within Human NFkB p100/NFKB2. The exact sequence is proprietary.
    Database link: Q00653

  • Positive control

    • WB: Jurkat, HeLa, Daudi, HCT116 and MCF7 cell lysates. ICC/IF: Wild-type HAP1 cells.
  • General notes

     This product was previously labelled as NFkB p100

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR4686-66
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • NFkB
    • p52
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • NFkB
    • p52

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab214265)
    • Alexa Fluor® 647 Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab214713)
    • Anti-NFkB p100/NFKB2 antibody [EPR4686-66] - BSA and Azide free (ab249847)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell line (ab262323)
    • Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (ab266883)
  • KO cell lysates

    • Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell lysate (ab257245)
    • Human NFKB2 (NFkB p100/NFKB2) knockout Hep G2 cell lysate (ab257247)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • MCF7 whole cell lysate (ab29537)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Daudi whole cell lysate (ab3951)
    • Jurkat whole cell lysate (ab7899)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab175192 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF
1/250.
IP
1/10 - 1/100.
WB (1)
1/10000 - 1/50000. Predicted molecular weight: 97 kDa.
Notes
ICC/IF
1/250.
IP
1/10 - 1/100.
WB
1/10000 - 1/50000. Predicted molecular weight: 97 kDa.
  • Application notes
    Is unsuitable for Flow Cyt or IHC-P.
  • Target

    • Relevance

      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 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. In a non-canonical activation pathway, the MAP3K14-activated CHUK/IKKA homodimer phosphorylates NFKB2/p100 associated with RelB, inducing its proteolytic processing to NFKB2/p52 and the formation of NF-kappa-B RelB-p52 complexes. The NF-kappa-B heterodimeric RelB-p52 complex is a transcriptional activator. The NF-kappa-B p52-p52 homodimer is a transcriptional repressor. NFKB2 appears to have dual functions such as cytoplasmic retention of attached NF-kappa-B proteins by p100 and generation of p52 by a cotranslational processing. The proteasome-mediated process ensures the production of both p52 and p100 and preserves their independent function. p52 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. p52 and p100 are respectively the minor and major form; the processing of p100 being relatively poor. Isoform p49 is a subunit of the NF-kappa-B protein complex, which stimulates the HIV enhancer in synergy with p65. In concert with RELB, regulates the circadian clock by repressing the transcriptional activator activity of the CLOCK-ARNTL/BMAL1 heterodimer.
    • Cellular localization

      Cytoplasmic and Nuclear
    • Database links

      • Entrez Gene: 4791 Human
      • Entrez Gene: 18034 Mouse
      • Entrez Gene: 309452 Rat
      • Omim: 164012 Human
      • SwissProt: Q00653 Human
      • SwissProt: Q9WTK5 Mouse
      • Unigene: 73090 Human
      • Unigene: 102365 Mouse
      • Unigene: 204814 Rat
      see all
    • Alternative names

      • 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
      • Oncogene Lyt 10 antibody
      • p100 antibody
      • Transcription factor NFKB2 antibody
      see all

    Images

    • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: NFκB p100 knockout HAP1 cell lysate (20 µg)
      Lane 3: Jurkat cell lysate (20 µg)
      Lane 4: HeLa cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab175192 observed at 97 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab175192 was shown to specifically react with NFκB p100 when NFκB p100 knockout samples were used. Wild-type and NFκB p100 knockout samples were subjected to SDS-PAGE. ab175192 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

      ab175192 staining NFkB p100/NFKB2 in wild-type HAP1 cells (top panel) and NFkB p100/NFKB2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab175192 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/1000 dilution

      Lane 1 : Wild-type HCT116 cell lysate
      Lane 2 : NFKB2 CRISPR/Cas9 edited HCT116 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 97 kDa
      Observed band size: 120 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab175192 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266883 (CRISPR/Cas9 edited cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 CRISPR/Cas9 edited HCT117 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/1000 dilution

      Lane 1 : Wild-type HepG2 cell lysate
      Lane 2 : NFKB2 CRISPR/Cas9 edited HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 97 kDa
      Observed band size: 120 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab175192 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HepG2 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab262323 (CRISPR/Cas9 edited cell lysate ab257247) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HepG2 and NFKB2 CRISPR/Cas9 edited HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : Daudi cell lysates
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 97 kDa

    • Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Immunoprecipitation - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

      Western blot analysis on immunoprecipitation pellet from Jurkat cell lysate labeling NFkB p100/NFKB2 with ab175192 at 1/10 dilution.

    • Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)
      Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (ab175192)

    Protocols

    • Western blot protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab175192? Please let us know so that we can cite the reference in this datasheet.

    ab175192 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Western blot abreview for Anti-NFkB p100/NFKB2 antibody [EPR4686-66]

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    Abreviews
    Abreviews
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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    50000 cells
    Specification
    293T
    Blocking step
    Milk as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C
    Read More

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    Submitted Aug 14 2019

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