Name: Anti-IKK alpha + IKK beta antibody [EPR16628]
Brand: RabMAb
SKU: ab178870
Rating: 5 (2 reviews)

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

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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.

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

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16628
Isotype

IgG
Research areas

Alternative Versions
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit (ab102918)
- APC Conjugation Kit (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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.

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
- Entrez Gene: 1147 Human
- Entrez Gene: 3551 Human
- Entrez Gene: 12675 Mouse
- Entrez Gene: 16150 Mouse
- Omim: 600664 Human
- Omim: 603258 Human
- SwissProt: O14920 Human
- SwissProt: O15111 Human
- SwissProt: O88351 Mouse
- SwissProt: Q60680 Mouse
- SwissProt: Q9QY78 Rat
- Unigene: 198998 Human
- Unigene: 413513 Human
see all
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

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

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

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

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

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

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

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

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

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.
Immunocytochemistry/ Immunofluorescence - Anti-IKK alpha + IKK beta [EPR16628] antibody (ab178870)

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

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

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

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

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

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.

Flow cytometry protocols
Immunoprecipitation protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

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

Publishing research using ab178870? Please let us know so that we can cite the reference in this datasheet.

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

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This product has been referenced in:

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