Recombinant
RabMAb

Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870)

Rabbit recombinant monoclonal IKK alpha + IKK beta antibody [EPR16628]. Validated in WB, IP, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 6 publication(s).

Overview

  • Product name
    Anti-IKK alpha + IKK beta antibody [EPR16628]
    See all IKK alpha + IKK beta primary antibodies
  • Description
    Rabbit monoclonal [EPR16628] to IKK alpha + IKK beta
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, ICC/IF, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human IKK alpha + IKK beta aa 450 to the C-terminus. The exact sequence is proprietary.
    Database link: O14920

  • Positive control
    • WB: HeLa, Daudi, HL-60, A431, 293T, C6, Raw264.7 and NIH/3T3 whole cell lysates. Human fetal kidney lysates. Mouse brain, Mouse kidney, Mouse spleen and Rat kidney lysates. ICC/IF: HeLa cells. IP: Jurkat whole cell extract.
  • 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.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178870 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
IP 1/40.
ICC/IF 1/50.
WB 1/1000. Detects a band of approximately 75, 87 kDa (predicted molecular weight: 85, 87 kDa).
Flow Cyt Use at an assay dependent concentration.

Target

  • Relevance
    IKK is a serine protein kinase, and the IKK complex contains alpha and beta subunits (IKK alpha and IKK beta). IKK alpha and IKK beta interact with each other and both are essential for NFkB activation.
  • Cellular localization
    Cytoplasmic and Nuclear; Membrane raft. Note: IKK beta is colocalized with DPP4 in membrane rafts.
  • Database links
  • Alternative names
    • CHUK antibody
    • Conserved helix loop helix ubiquitous kinase antibody
    • I kappa B kinase 1 antibody
    • I kappa B kinase 2 antibody
    • I Kappa B kinase alpha antibody
    • I Kappa B kinase beta antibody
    • IkB kinase alpha subunit antibody
    • IkBKA antibody
    • IkBKB antibody
    • IKK a kinase antibody
    • IKK alpha antibody
    • IKK beta antibody
    • IKK-alpha antibody
    • IKK-beta antibody
    • IKK1 antibody
    • IKK2 antibody
    • IKKA antibody
    • IKKB antibody
    • IMD15 antibody
    • Inhibitor of kappa light polypeptide gene enhancer in B cells kinase beta antibody
    • Inhibitor of nuclear factor kappa-B kinase subunit alpha antibody
    • Inhibitor of nuclear factor kappa-B kinase subunit beta antibody
    • NFKBIKA antibody
    • NFKBIKB antibody
    • Nuclear factor NF kappa B inhibitor kinase beta antibody
    • Nuclear factor NFkappaB inhibitor kinase alpha antibody
    • TCF 16 antibody
    • TCF16 antibody
    • Transcription factor 16 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling IKK alpha + IKK beta with ab178870 at 1/50 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody (ab150077) at 1/200 dilution (green). Nuclear and cytoplasm staining is detected. 

    Tubulin is detected with Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). The nuclear counter stain is DAPI (blue). 

     

    The two negative controls were :-

    -ve control 1 - ab178870 at 1/50 diltuion followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
    -ve control 2 - Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) at 1/200 dilution.

  • All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates
    Lane 3 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa
    why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.

     

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IKK alpha + IKK beta with purified ab178870 at 1/130 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

  • All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

    Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates
    Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.


    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution + Human fetal kidney lysates at 20 µg

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

    Predicted band size: 85, 87 kDa
    Observed band size: 87 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IKK alpha + IKK beta antibody [EPR16628] (ab178870) at 1/1000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse kidney lysates
    Lane 3 : Mouse spleen lysates
    Lane 4 : Rat kidney lysates
    Lane 5 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 7 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa why is the actual band size different from the predicted?



    Blocking/dilution buffer: 5% NFDM/TBST.


    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab178870 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST. 

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

     

     

  • Cross-Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab32041 (IKK alpha) at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST.

  • This immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% triton-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells is a comparison between ab178870 and a competitor’s leading rabbit polyclonal antibody.

    Labeling of IKK alpha + IKK beta with ab178870 is conducted at 1/50 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) secondary antibody (ab150077) at 1/200 dilution.
    Labeling of IKK alpha + IKK beta with the competitor antibody is conducted at 1/40 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor ® 488) at 1/200 dilution.

    Tubulin is detected with Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/400 dilution (red). The nuclear counter stain is DAPI (blue).
    The two negative controls were :
    -ve control 1 - ab178870 at 1/50 diltuion followed by Goat anti mouse IgG (Alexa Fluor®594) at 1/400 dilution.
    -ve control 2 - Mouse anti-Tubulin antibody (ab7291) at 1/500 dilution followed by Goat anti rabbit IgG (Alexa Fluor®488) at 1/200 dilution.

  • All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates
    Lane 3 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa why is the actual band size different from the predicted?



    This western blot image is a comparison of ab178870 against a competitor's leading rabbit polyclonal antibody.

    Blocking/dilution buffer: 5% NFDM/TBST.

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200

    Lane 1 : A431 (Human epidermoid carcinoma) whole cell lysates
    Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa why is the actual band size different from the predicted?



    This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

    Blocking/dilution buffer: 5% NFDM/TBST.

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution + Human fetal kidney lysates

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

    Predicted band size: 85, 87 kDa
    Observed band size: 87 kDa why is the actual band size different from the predicted?



    This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

    Blocking/dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870) at 1/1000 dilution, and a competitor's Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/200 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse kidney lysates
    Lane 3 : Mouse spleen lysates
    Lane 4 : Rat kidney lysates
    Lane 5 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 7 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

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

    Predicted band size: 85, 87 kDa
    Observed band size: 75,87 kDa why is the actual band size different from the predicted?



    This western blot image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

    Blocking/dilution buffer: 5% NFDM/TBST.

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

  • This immunoprecipation image is a comparison between ab178870 and a competitor's leading rabbit polyclonal antibody.

    Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using ab178870 at 1/40 dilution. Western blot detection was performed using ab178870 at 1/1000 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution. The blocking and diluting buffer was 5% NFDM/TBST.

    Immunoprecipitation of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell extracts using competitor’s Anti-IKK alpha + IKK beta rabbit polyclonal antibody at 1/12 dilution. Western blot detection was performed using competitor antibody at 1/100 dilution followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/100 dilution. The blocking and diluting buffer was 5% NFDM/TBST. 

    ab178870 could recognize 3 isoforms of IKK beta with the MWs of 87kDa, 86kDa and 80kDa, respectively. It could also recognize IKK alpha.

     

References

This product has been referenced in:
  • Zhu S  et al. Tanshinone-IIA attenuates the deleterious effects of oxidative stress in osteoporosis through the NF-?B signaling pathway. Mol Med Rep 17:6969-6976 (2018). Read more (PubMed: 29568934) »
  • Yang T  et al. lncRNA-NKILA/NF-?B feedback loop modulates laryngeal cancer cell proliferation, invasion, and radioresistance. Cancer Med 7:2048-2063 (2018). WB ; Human . Read more (PubMed: 29573243) »
See all 6 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A375 (melanoma cell line))
Permeabilization
Yes - Triton 100x 0,5%
Specification
A375 (melanoma cell line)
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Paraformaldehyde

Cristina Penas Lago

Verified customer

Submitted Dec 11 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (mesenchymal stem cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
mesenchymal stem cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Kate Hawkins

Verified customer

Submitted Mar 22 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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