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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-NFkB p100/NFKB2 抗体 [EPR4686] - BSA and Azide free (ab174482)

Submit a review Submit a question References (1)
Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
  • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
  • Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)

Key features and details

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

Conjugates logo Related conjugates and formulations

Unconjugated

製品の概要

  • 製品名

    Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free
    NFkB p100/NFKB2 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR4686] to NFkB p100/NFKB2 - BSA and Azide free

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, ICC/IFmore details
    適用なし: Flow Cyt or IHC-P

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse

  • 免疫原

    Synthetic peptide within Human NFkB p100/NFKB2 aa 700 to the C-terminus. The exact sequence is proprietary.
    Database link: Q00653

  • ポジティブ･コントロール

    • WB: Jurkat, HeLa, ECV-304, HepG2, HCT116 and MCF7 cell lysates ICC/IF: Wild-type HAP1 cells.

  • 特記事項

    ab174482 is the carrier-free version of ab109440.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab174482の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 97 kDa).
ICC/IF
Use at an assay dependent concentration.
特記事項
WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 97 kDa).
ICC/IF
Use at an assay dependent concentration.
追加情報
Is unsuitable for Flow Cyt or IHC-P.

ターゲット情報

  • 関連性

    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.

  • 細胞内局在

    Cytoplasmic and Nuclear

  • 参照データベース

    • 別名

      • 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

    画像

    • Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)

      This data was developed using ab109440, the same antibody clone in a different buffer formulation.

      Lane 1 Wild-type HAP1 cell lysate (20 µg)

      Lane 2 NF?B 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 - ab109440 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab109440 was shown to specifically react with NF?B p100 when NF?B p100 knockout samples were used. Wild-type and NFB p100 knockout samples were subjected to SDS-PAGE. ab109440 and ab109440 (loading control to GAPDH) were diluted 1/10000 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] - BSA and Azide free (ab174482)
      Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      This data was developed using ab109440, the same antibody clone in a different buffer formulation.ab109440 staining NFkB p100/p52 in wild-type HAP1 cells (top panel) and NFkB p100/p52 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 ab109440 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] - BSA and Azide free (ab174482)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) 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?



      This data was developed using the same antibody clone in a different buffer formulation (ab109440).

      Lanes 1- 2: Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109440 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 HCT116 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. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 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] - BSA and Azide free (ab174482)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) 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?



      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109440).

      Lanes 1- 2: Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab109440 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. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 1000 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] - BSA and Azide free (ab174482)
      Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      All lanes : Anti-NFkB p100/NFKB2 antibody [EPR4686] (ab109440) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate
      Lane 3 : ECV-304 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 97 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?



      This data was developed using ab109440, the same antibody clone in a different buffer formulation.

       

    • Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)
      Anti-NFkB p100/NFKB2 antibody [EPR4686] - BSA and Azide free (ab174482)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (1)

    ab174482 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab174482 は 1 報の論文で使用されています。

    • Liu D  et al. ASPP1 deficiency promotes epithelial-mesenchymal transition, invasion and metastasis in colorectal cancer. Cell Death Dis 11:224 (2020). PubMed: 32269211

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