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  1. Link

    ikk-alpha-antibody-y463-ab32041.pdf

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

Recombinant Anti-IKK alpha antibody [Y463] (ab32041)

  • Datasheet
  • SDS
Reviews (6)Q&A (3)References (53)

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Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
  • Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
  • Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
  • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)
  • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
  • Immunoprecipitation - Anti-IKK alpha antibody [Y463] (ab32041)
  • Anti-IKK alpha antibody [Y463] (ab32041)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y463] to IKK alpha
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP

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Overview

  • Product name

    Anti-IKK alpha antibody [Y463]
    See all IKK alpha primary antibodies
  • Description

    Rabbit monoclonal [Y463] to IKK alpha
  • Host species

    Rabbit
  • Specificity

    The rat recommendation is based on the WB results. We do not guarantee IHC-P for rat.

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1-WT; Daudi,RAW 264.7 and C6 whole cell lysate. Mouse and rat kidney lysate. Flow Cyt (intra): HAP1-WT and Daudi cells; IP: HeLa cell lysate; IHC-P: Human ovarian cancer and mouse kidney tissue.
  • 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.

    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.

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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y463
  • 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
    • 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 alpha antibody [Y463] - BSA and Azide free (ab169743)
    • Alexa Fluor® 488 Anti-IKK alpha antibody [Y463] (ab200412)
    • Alexa Fluor® 647 Anti-IKK alpha antibody [Y463] (ab200414)
    • HRP Anti-IKK alpha antibody [Y463] (ab200415)
    • APC Anti-IKK alpha antibody [Y463] (ab221910)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Daudi whole cell lysate (ab3951)
  • Recombinant Protein

    • Recombinant human IKK alpha protein (Active) (ab102103)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32041 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
Flow Cyt (Intra)
1/20.

For unpurified use at 1/50.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB (4)
1/1000. Detects a band of approximately 88 kDa (predicted molecular weight: 85 kDa).

For unpurified use at 1/10000 - 1/50000.

IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The rat recommendation is based on the WB results. We do not guarantee IHC-P for rat.

IP (1)
1/60.
Notes
Flow Cyt (Intra)
1/20.

For unpurified use at 1/50.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/1000. Detects a band of approximately 88 kDa (predicted molecular weight: 85 kDa).

For unpurified use at 1/10000 - 1/50000.

IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

The rat recommendation is based on the WB results. We do not guarantee IHC-P for rat.

IP
1/60.

Target

  • Function

    Acts as part of the IKK complex in the conventional pathway of NF-kappa-B activation and 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. As part of the non-canonical pathway of NF-kappa-B activation, 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. Also phosphorylates NCOA3. Phosphorylates 'Ser-10' of histone H3 at NF-kappa-B-regulated promoters during inflammatory responses triggered by cytokines.
  • Tissue specificity

    Widely expressed.
  • Involvement in disease

    Defects in CHUK are the cause of cocoon syndrome (COCOS) [MIM:613630]; also known as fetal encasement syndrome. COCOS is a lethal syndrome characterized by multiple fetal malformations including defective face and seemingly absent limbs, which are bound to the trunk and encased under the skin.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Phosphorylated by MAP3K14/NIK, AKT and to a lesser extent by MEKK1, and dephosphorylated by PP2A. Autophosphorylated.
    Acetylation of Thr-179 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B signaling pathway.
  • Cellular localization

    Cytoplasm. Nucleus. Shuttles between the cytoplasm and the nucleus.
  • Target information above from: UniProt accession O15111 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1147 Human
    • Entrez Gene: 309361 Rat
    • Omim: 600664 Human
    • SwissProt: O15111 Human
    • SwissProt: Q60680 Mouse
    • Unigene: 198998 Human
    • Unigene: 3996 Mouse
    • Alternative names

      • chuk antibody
      • CHUK1 antibody
      • Conserved Helix Loop Helix Ubiquitous Kinase antibody
      • Conserved helix loop ubiquitous kinase antibody
      • Conserved helix-loop-helix ubiquitous kinase antibody
      • I Kappa B Kinase 1 antibody
      • I Kappa B Kinase Alpha antibody
      • I-kappa-B kinase 1 antibody
      • I-kappa-B kinase alpha antibody
      • IkappaB kinase antibody
      • IkB kinase alpha subunit antibody
      • IkBKA antibody
      • IKK 1 antibody
      • IKK A antibody
      • IKK a kinase antibody
      • IKK-A antibody
      • IKK-alpha antibody
      • IKK1 antibody
      • IKKA antibody
      • IKKA_HUMAN antibody
      • Inhibitor Of Kappa Light Polypeptide Gene Enhancer In B Cells antibody
      • Inhibitor Of Nuclear Factor Kappa B Kinase Alpha Subunit antibody
      • Inhibitor of nuclear factor kappa-B kinase subunit alpha antibody
      • NFKBIKA antibody
      • Nuclear Factor Kappa B Inhibitor Kinase Alpha antibody
      • Nuclear factor NF kappa B inhibitor kinase alpha antibody
      • Nuclear factor NF-kappa-B inhibitor kinase alpha antibody
      • Nuclear factor NFkappaB inhibitor kinase alpha antibody
      • Nuclear Factor Of Kappa Light Chain Gene Enhancer In B Cells Inhibitor antibody
      • TCF-16 antibody
      • TCF16 antibody
      • Transcription factor 16 antibody
      see all

    Images

    • Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
      Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
      All lanes : Anti-IKK alpha antibody [Y463] (ab32041) at 1/1000 dilution (Purified)

      Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 2 : Mouse kidney lysate
      Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysate
      Lane 4 : Rat kidney lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 85 kDa

    • Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
      Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
      Anti-IKK alpha antibody [Y463] (ab32041) at 1/10000 dilution + Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

      Predicted band size: 85 kDa

    • Western blot - Anti-IKK alpha antibody [Y463] (ab32041)
      Western blot - Anti-IKK alpha antibody [Y463] (ab32041)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: IKK alpha knockout HAP1 cell lysate (20 µg)
      Lanes 1 and 2: Merged signal (red and green). Green - ab32041 observed at 84 kDa. Red - loading control, ab8226, observed at 42 kDa.

      ab32041 (Unpurified format) was shown to specifically react with IKK alpha when IKK alpha knockout samples were used. Wild-type and IKK alpha knockout samples were subjected to SDS-PAGE. ab32041 and ab8226 (loading control to beta actin) were diluted 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian cancer tissue sections labeling IKK alpha with purified ab32041 at 1/50 dilution (3.92 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
    • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)
      Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)

      Overlay histogram showing HAP1 wildtype (green line) and HAP1-CHUK knockout cells (red line) stained with ab32041. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab32041, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CHUK knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions. 

       

       

    • Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)
      Flow Cytometry (Intracellular) - Anti-IKK alpha antibody [Y463] (ab32041)

      Intracellular Flow Cytometry analysis of Daudi (Human Burkitt's lymphoma lymphoblast) cells labelling IKK alpha with purified ab32041 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling IKK alpha with purified ab32041 at 1/50 dilution (3.92 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
    • Immunoprecipitation - Anti-IKK alpha antibody [Y463] (ab32041)
      Immunoprecipitation - Anti-IKK alpha antibody [Y463] (ab32041)

      Purified ab32041 at 1/60 dilution (2µg) immunoprecipitating IKK alpha in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab32041 + HeLa whole cell lysate.
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab169743 in HeLa whole cell lysate.
      VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 88 kDa

    • Anti-IKK alpha antibody [Y463] (ab32041)
      Anti-IKK alpha antibody [Y463] (ab32041)

    Protocols

    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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (53)

    Publishing research using ab32041? Please let us know so that we can cite the reference in this datasheet.

    ab32041 has been referenced in 53 publications.

    • Zhang WB  et al. Protective Effects of Oroxylin A against Doxorubicin-Induced Cardiotoxicity via the Activation of Sirt1 in Mice. Oxid Med Cell Longev 2021:6610543 (2021). PubMed: 33542782
    • Tian S  et al. Curcumin inhibits the growth of liver cancer by impairing myeloid-derived suppressor cells in murine tumor tissues. Oncol Lett 21:286 (2021). PubMed: 33732362
    • Li Q  et al. Carminic acid supplementation protects against fructose-induced kidney injury mainly through suppressing inflammation and oxidative stress via improving Nrf-2 signaling. Aging (Albany NY) 13:10326-10353 (2021). PubMed: 33819919
    • Lu C  et al. Tumor necrosis factor receptor type 1-associated death domain (TRADD) regulates epithelial-mesenchymal transition (EMT), M1/M2 macrophage polarization and ectopic endometrial cysts formation in endometriosis. Ann Transl Med 9:148 (2021). PubMed: 33569450
    • Xu Y  et al. Identification of the Key Role of NF-?B Signaling Pathway in the Treatment of Osteoarthritis With Bushen Zhuangjin Decoction, a Verification Based on Network Pharmacology Approach. Front Pharmacol 12:637273 (2021). PubMed: 33912052
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-6 of 6 Abreviews

    Western blot abreview for Anti-IKK alpha antibody [Y463]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Monkey Tissue lysate - whole (Prefrontal lobe)
    Gel Running Conditions
    Reduced Denaturing (gel 10%)
    Loading amount
    20 µg
    Specification
    Prefrontal lobe
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 07 2021

    Immunoprecipitation abreview for Anti-IKK alpha antibody [Y463]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (Raw 264.7)
    Total protein in input
    100 µg
    Specification
    Raw 264.7
    Treatment
    2 hrs LPS
    Immuno-precipitation step
    Protein G
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jul 31 2012

    Western blot abreview for Anti-IKK alpha antibody [Y463]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Raw 264.7)
    Loading amount
    50 µg
    Specification
    Raw 264.7
    Treatment
    2 hrs LPS
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 08 2012

    Western blot abreview for Anti-IKK alpha antibody [Y463]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Guinea pig Tissue lysate - whole (myometrium pregnant and non-pregnant)
    Loading amount
    30 µg
    Specification
    myometrium pregnant and non-pregnant
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 14 2011

    Western blot abreview for Anti-IKK alpha antibody [Y463]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Epithelial, Neutrophils, monocytes)
    Loading amount
    500000 cells
    Specification
    Epithelial, Neutrophils, monocytes
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Sep 02 2009

    Immunohistochemistry (Frozen sections) abreview for Anti-IKK alpha antibody [Y463]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skin)
    Specification
    skin
    Fixative
    Paraformaldehyde
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6%
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Apr 24 2007

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