Validated using a knockout cell line
Recombinant
RabMAb

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

Overview

  • Product name

    Anti-NFkB p105 / p50 antibody [E381] (HRP)
    See all NFkB p105 / p50 primary antibodies
  • Description

    Rabbit monoclonal [E381] to NFkB p105 / p50 (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

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

    Synthetic peptide within NFkB p105/ p50 aa 300-400. The exact sequence is proprietary.

  • Positive control

    • WB: HeLa whole cell lysate.
  • General notes

    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

Applications

Our Abpromise guarantee covers the use of ab195854 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 1/2000. Detects a band of approximately 50, 105 kDa (predicted molecular weight: 50 kDa).

Target

  • Function

    NF-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 similarities

    Contains 7 ANK repeats.
    Contains 1 death domain.
    Contains 1 RHD (Rel-like) domain.
  • Domain

    The 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 localization

    Nucleus. 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 antibody
    • EBP 1 antibody
    • EBP-1 antibody
    • EBP1 antibody
    • KBF1 antibody
    • MGC54151 antibody
    • nf b antibody
    • NF kappa B antibody
    • NF kappaB antibody
    • NF kappabeta antibody
    • NF kB1 antibody
    • NF-kappaB antibody
    • NFkappaB antibody
    • NFKB 1 antibody
    • NFKB p105 antibody
    • NFKB p50 antibody
    • NFKB-p105 antibody
    • Nfkb1 antibody
    • NFKB1_HUMAN antibody
    • Nuclear factor kappa B DNA binding subunit antibody
    • Nuclear factor kappa-B 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 chain gene enhancer in B cells 1 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
    • Transcription factor NFKB1 antibody
    see all

Images

  • All lanes : Anti-NFkB p105 / p50 antibody [E381] (HRP) (ab195854) at 1/2000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : Nfkb1 (NFkB p105 / p50) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

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


    Exposure time: 20 minutes


    ab195854 was shown to recognize NFkB p105 / p50 in wild-type HAP1 cells as signal was lost at the expected MW in Nfkb1 (NFkB p105 / p50) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Nfkb1 (NFkB p105 / p50) knockout samples were subjected to SDS-PAGE. Ab195854 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Anti-NFkB p105 / p50 antibody [E381] (HRP) (ab195854) at 1/2000 dilution + HeLa whole cell lysate (ab150035) at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 20 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab195854 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

This product has been referenced in:

  • Qian Y  et al. Lactobacillus casei Strain Shirota Enhances the In Vitro Antiproliferative Effect of Geniposide in Human Oral Squamous Carcinoma HSC-3 Cells. Molecules 23:N/A (2018). Read more (PubMed: 29751513) »
See 1 Publication for this product

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