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

Recombinant Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (1)

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Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
  • Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
  • Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
  • Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
  • Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14660] to IKK gamma/NEMO
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Protein
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Recombinant Human IKK gamma/NEMO protein (ab125589)
Knockout
Product image
Human IKBKG (IKK gamma/NEMO) knockout HEK-293T cell line (ab266674)
Primary
Product image
Anti-COPS6 antibody (ab77299)

View more associated products

Overview

  • Product name

    Anti-IKK gamma/NEMO antibody [EPR14660]
    See all IKK gamma/NEMO primary antibodies
  • Description

    Rabbit monoclonal [EPR14660] to IKK gamma/NEMO
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T, HeLa, Jurkat, K572 and human fetal brain lysates. ICC/IF: Jurkat cells.
  • General notes

    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: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR14660
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • NFkB
    • IKK
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Inflammatory mediators
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • NFkB
    • IKK

Associated products

  • Alternative Versions

    • Anti-IKK gamma/NEMO antibody [EPR14660] - BSA and Azide free (ab250964)
  • 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 IKBKG (IKK gamma/NEMO) knockout HEK-293T cell line (ab266674)
  • KO cell lysates

    • Human IKBKG (IKK gamma/NEMO) knockout HEK-293T cell lysate (ab257153)
  • Positive Controls

    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab7899)
    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human IKK gamma/NEMO protein (ab125589)
  • Related Products

    • Recombinant Human IKK gamma/NEMO protein (ab125589)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab188569 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
1/10000 - 1/50000. Predicted molecular weight: 48 kDa.
ICC/IF
1/100.
Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 48 kDa.
ICC/IF
1/100.

Target

  • Function

    Regulatory subunit of the IKK core complex which phosphorylates inhibitors of NF-kappa-B thus leading to the dissociation of the inhibitor/NF-kappa-B complex and ultimately the degradation of the inhibitor. Also considered to be a mediator for TAX activation of NF-kappa-B. Could be implicated in NF-kappa-B-mediated protection from cytokine toxicity (By similarity). Essential for viral activation of IRF3.
  • Tissue specificity

    Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
  • Involvement in disease

    Defects in IKBKG are the cause of ectodermal dysplasia anhidrotic with immunodeficiency X-linked (EDAID) [MIM:300291]; also known as hypohidrotic ectodermal dysplasia with immunodeficiency (HED-ID). Is a form of ectoderma dysplasia, a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. Characterized by absence of sweat glands, sparse scalp hair, rare conical teeth and immunological abnormalities resulting in severe infectious diseases.
    Defects in IKBKG are the cause of ectodermal dysplasia anhidrotic with immunodeficiency-osteopetrosis-lymphedema (OLEDAID) [MIM:300301].
    Defects in IKBKG are a cause of immunodeficiency NEMO-related without anhidrotic ectodermal dysplasia (NEMOID) [MIM:300584]; also called immunodeficiency without anhidrotic ectodermal dysplasia, isolated immunodeficiency or pure immunodeficiency. Patients manifest immunodeficiency not associated with other abnormalities, and resulting in increased infection susceptibility. Patients suffer from multiple episodes of infectious diseases.
    Defects in IKBKG are the cause of susceptibility to X-linked familial atypical micobacteriosis type 1 (AMCBX1) [MIM:300636]; also known as X-linked disseminated atypical mycobacterial infection type 1 or X-linked susceptibility to mycobacterial disease type 1. AMCBX1 is the X-linked recessive form of mendelian susceptibility to mycobacterial disease (MSMD). MSMD is a congenital syndrome resulting in predisposition to clinical disease caused by weakly virulent mycobacterial species, such as bacillus Calmette-Guerin vaccines and non-tuberculous, environmental mycobacteria. Patients are also susceptible to the more virulent species Mycobacterium tuberculosis.
    Defects in IKBKG are the cause of recurrent isolated invasive pneumococcal disease type 2 (IPD2) [MIM:300640]. Recurrent invasive pneumococcal disease (IPD) is defined as two episodes of IPD occurring at least 1 month apart, whether caused by the same or different serotypes or strains. Recurrent IPD occurs in at least 2% of patients in most series, making IPD the most important known risk factor for subsequent IPD.
    Defects in IKBKG are the cause of incontinentia pigmenti (IP) [MIM:308300]; formerly designed familial incontinentia pigmenti type II (IP2). IP is a genodermatosis usually prenatally lethal in males. In affected females, it causes abnormalities of the skin, hair, eyes, nails, teeth, skeleton, heart, and central nervous system. The prominent skin signs occur in four classic cutaneous stages: perinatal inflammatory vesicles, verrucous patches, a distinctive pattern of hyperpigmentation and dermal scarring.
  • Sequence similarities

    Contains 1 C2HC-type zinc finger.
  • Domain

    The leucine-zipper domain and the C2HC-type zinc-finger are essential for polyubiquitin binding and for the activation of IRF3.
  • Post-translational
    modifications

    Phosphorylation at Ser-68 attenuates aminoterminal homodimerization.
    Polyubiquitinated on Lys-285 through 'Lys-63'; the ubiquitination is mediated by NOD2 and RIPK2 and probably plays a role in signaling by facilitating interactions with ubiquitin domain-containing proteins and activates the NF-kappa-B pathway. Polyubiquitinated on Lys-399 through 'Lys-63'; the ubiquitination is mediated by BCL10, MALT1 and TRAF6 and probably plays a role in signaling by facilitating interactions with ubiquitin domain-containing proteins and activates the NF-kappa-B pathway. Monoubiquitinated on Lys-277 and Lys-309; promotes nuclear export. Linear polyubiquitinated on Lys-285; the head-to-tail polyubiquitination is mediated by the LUBAC complex. Linear polyubiquitinated on Lys-309; the head-to-tail polyubiquitination is mediated by the LUBAC complex.
    Sumoylated on Lys-277 and Lys-309 by SUMO1; the modification results in phosphorylation of Ser-85 by ATM leading to a replacement of the sumoylation by mono-ubiquitination on these residues.
  • Cellular localization

    Cytoplasm. Nucleus. Sumoylated NEMO accumulates in the nucleus in response to genotoxic stress.
  • Target information above from: UniProt accession Q9Y6K9 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8517 Human
    • Omim: 300248 Human
    • SwissProt: Q9Y6K9 Human
    • Unigene: 43505 Human
    • Alternative names

      • IkB kinase associated protein 1 antibody
      • IkB kinase subunit gamma antibody
      • Inhibitor of nuclear factor kappa B kinase subunit gamma antibody
      • AMCBX1 antibody
      • FIP 3 antibody
      • FIP-3 antibody
      • FIP3 antibody
      • Fip3p antibody
      • I kappa B kinase gamma antibody
      • I-kappa-B kinase subunit gamma antibody
      • IkB kinase gamma subunit antibody
      • IkB kinase subunit gamma antibody
      • IkB kinase-associated protein 1 antibody
      • Ikbkg antibody
      • IKK gamma antibody
      • IKK-gamma antibody
      • IKKAP1 antibody
      • IKKG antibody
      • IMD33 antibody
      • Incontinentia pigmenti antibody
      • Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase gamma antibody
      • Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase of, gamma antibody
      • Inhibitor of nuclear factor kappa-B kinase subunit gamma antibody
      • IP antibody
      • IP1 antibody
      • IP2 antibody
      • IPD2 antibody
      • NEMO antibody
      • NEMO_HUMAN antibody
      • NF kappa B essential modifier antibody
      • NF kappa B essential modulator antibody
      • NF-kappa-B essential modifier antibody
      • NF-kappa-B essential modulator antibody
      • ZC2HC9 antibody
      see all

    Images

    • Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IKK gamma/NEMO knockout HAP1 cell lysate (20 µg)
      Lane 3: Jurkat cell lysate (20 µg)

      Lanes 1 - 3: Merged signal (red and green). Green - ab188569 observed at 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab188569 was shown to react with IKK gamma/NEMO in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when IKK gamma/NEMO knockout samples were examined. Wild-type and IKK gamma/NEMO knockout samples were subjected to SDS-PAGE. ab188569 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/1000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      Immunocytochemistry/ Immunofluorescence - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)

      Immunocytochemical analysis of Jurkat cells fixed in 4% paraformaldehyde labeling IKK gamma/NEMO with ab188569 at 1/100 dilution and Goat anti rabbit IgG(Alexa Fluor® 555) at 1/200 dilution.  Counterstained with DAPI.

    • Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      All lanes : Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : IKBKG CRISPR/Cas9 edited HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 48 kDa
      Observed band size: 48 kDa



      Lanes 1- 2: Merged signal (red and green). Green - ab188569 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

       ab188569 was shown to react with IKK gamma/NEMO in wild-type HEK-293T cells in western blot. The band observed in CRISPR/Cas9 edited  cell line ab266674 (CRISPR/Cas9 edited cell lysate ab257153) lane below 48kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HEK-293T and IKBKG CRISPR/Cas9 edited HEK-293T 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. ab188569 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-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      Western blot - Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      All lanes : Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569) at 1/10000 dilution

      Lane 1 : Hela cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : Human fetal brain lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 48 kDa
      Additional bands at: 41 kDa (possible cleavage fragment)

    • Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)
      Anti-IKK gamma/NEMO antibody [EPR14660] (ab188569)

    Protocols

    • Western blot protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab188569? Please let us know so that we can cite the reference in this datasheet.

    ab188569 has been referenced in 1 publication.

    • Chen S  et al. Feline Infectious Peritonitis Virus Nsp5 Inhibits Type I Interferon Production by Cleaving NEMO at Multiple Sites. Viruses 12:N/A (2019). PubMed: 31905881

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