RabMAb

Anti-NFkB p105 / p50 antibody [E381] (ab32360)

Overview

  • Product nameAnti-NFkB p105 / p50 antibody [E381]
    See all NFkB p105 / p50 primary antibodies
  • Description
    Rabbit monoclonal [E381] to NFkB p105 / p50
  • SpecificityThis antibody will detect both forms: p50 and p105.
  • Tested applicationsWB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to NFkB p105/ p50 aa 300-400.

  • Positive control
    • WB: HeLa, MCF-7 and PC-12 cell lysates. IHC-P: human prostate carcinoma and bladder carcinoma tissues.
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions:

    Anti-NFkB p105 / p50 antibody (Alexa Fluor® 647) [E381] (ab196519)

    Anti-NFkB p105 / p50 antibody (HRP) [E381] (ab195854)

Properties

Applications

Our Abpromise guarantee covers the use of ab32360 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
WB 1/1000 - 1/10000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).
IHC-P 1/250.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/250 - 1/500.

  • Application notesIs unsuitable for Flow Cyt or ICC/IF.
  • Target

    • FunctionNF-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.
    • Sequence similaritiesContains 7 ANK repeats.
      Contains 1 death domain.
      Contains 1 RHD (Rel-like) domain.
    • DomainThe 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
      modifications
      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.
    • Cellular localizationNucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor.
    • Information by UniProt
    • Database links
    • Alternative names
      • DKFZp686C01211 antibody
      • DNA binding factor KBF1 antibody
      • DNA binding factor KBF1 EBP1 antibody
      • DNA binding factor KBF1 EBP1 antibody
      • DNA-binding factor KBF1 antibody
      • EBP 1 antibody
      • EBP-1 antibody
      • EBP1 antibody
      • KBF1 antibody
      • MGC54151 antibody
      • NF kappa B antibody
      • NF kappaB antibody
      • NF kappabeta antibody
      • NF kB1 antibody
      • NFkappaB antibody
      • NFKB 1 antibody
      • NFKB p105 antibody
      • NFKB p50 antibody
      • NFKB1 antibody
      • NFKB1_HUMAN antibody
      • Nuclear factor kappa B DNA binding subunit antibody
      • Nuclear factor kappa-B, subunit 1 antibody
      • Nuclear factor NF kappa B p105 subunit antibody
      • Nuclear factor NF kappa B p50 subunit antibody
      • Nuclear factor NF-kappa-B p50 subunit antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B cells 1 antibody
      • Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 antibody
      • p105 antibody
      • p50 antibody
      • p84/NF-kappa-B1 p98 antibody
      see all

    Anti-NFkB p105 / p50 antibody [E381] images

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified) + HeLa cell lysate at 20 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 50,105 kDa (why is the actual band size different from the predicted?)

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/50000 dilution (purified) + MCF-7 cell lysate at 10 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 105 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 50 kDa (possible non-specific binding).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/10000 dilution (purified)

      Lane 1 : PC-12 cell lysate
      Lane 2 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 50 kDa
      Observed band size : 105 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 50 kDa (possible non-specific binding).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-NFkB p105 / p50 antibody [E381] (ab32360) at 1/5000 dilution (unpurified) + HeLa cell lysate

      Predicted band size : 50 kDa
      Observed band size : 50,105 kDa (why is the actual band size different from the predicted?)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue labelling NFkB p105 / p50 with purified ab32360 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling NFkB p105 / p50 with unpurified ab32360 at a dilution of 1/250.

    Protocols

    References for Anti-NFkB p105 / p50 antibody [E381] (ab32360)

    This product has been referenced in:
    • Ma L  et al. Cluster of differentiation 166 (CD166) regulates cluster of differentiation (CD44) via NF-?B in liver cancer cell line Bel-7402. Biochem Biophys Res Commun 451:334-8 (2014). Read more (PubMed: 25094049) »
    • Li C  et al. Qishenyiqi protects ligation-induced left ventricular remodeling by attenuating inflammation and fibrosis via STAT3 and NF-?B signaling pathway. PLoS One 9:e104255 (2014). WB ; Rat . Read more (PubMed: 25122164) »

    See all 15 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (10%)
    Sample Rat Tissue lysate - whole (Skeletal muscle)
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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    Submitted Feb 20 2015

    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (10% gel)
    Sample Mouse Tissue lysate - whole (Skeletal muscle)
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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    Submitted Feb 20 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10mM Tris, 1 mM EDTA pH 9
    Sample Monkey Tissue sections (retina)
    Specification retina
    Permeabilization No
    Fixative Paraformaldehyde
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    Submitted Feb 10 2015

    Application Western blot
    Loading amount 200 µg
    Gel Running Conditions Reduced Denaturing (4-15% SDS-PAGE)
    Sample Monkey Tissue lysate - whole (macaque retina)
    Specification macaque retina
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Feb 06 2015

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step Dako Flex peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: High pH
    Sample Human Tissue sections (Tendon)
    Specification Tendon
    Permeabilization No
    Fixative 10% buffered formalin
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    Submitted Oct 22 2014

    Application Western blot
    Loading amount 25 µg
    Gel Running Conditions Reduced Denaturing (4–15%)
    Sample Mouse Tissue lysate - whole (Lung)
    Specification Lung
    Blocking step BSA as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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    Submitted Jul 01 2014

    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step hydrogen peroxide as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate, pH6.0, 100C, 20 min
    Sample Human Tissue sections (placenta)
    Specification placenta
    Permeabilization No
    Fixative Formaldehyde
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    Submitted Jul 01 2014

    Application Western blot
    Loading amount 50 µg
    Gel Running Conditions Reduced Denaturing (12% gel)
    Sample Mouse Tissue lysate - whole (Skeletal muscle)
    Specification Skeletal muscle
    Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

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    Verified customer

    Submitted Jan 09 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (spleen lymphoma)
    Specification spleen lymphoma
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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    Submitted Apr 09 2013

    Thank you for contacting us. Flow cytometry protocols are recommended below. For most of the antibodies we are not able to provide specific epitope information but it seems most targets are intracellular. Hence not on the membrane and cells need fi...

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