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Rabbit Recombinant Monoclonal IKK alpha antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 75 publications.


Images

Immunoprecipitation - Anti-IKK alpha antibody [Y463] (AB32041), expandable thumbnail
  • Western blot - Anti-IKK alpha antibody [Y463] (AB32041), expandable thumbnail
  • Western blot - Anti-IKK alpha antibody [Y463] (AB32041), expandable thumbnail
  • Western blot - Anti-IKK alpha antibody [Y463] (AB32041), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (AB32041), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Expected
Expected
Rat
Expected
Expected
Tested
Expected

Tested
Tested

Species
Human
Dilution info
1/50
Notes

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

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

Species
Mouse
Dilution info
1/50
Notes

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

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

Expected
Expected

Species
Rat
Dilution info
1/50
Notes

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

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

Tested
Tested

Species
Human
Dilution info
1/60
Notes

-

Expected
Expected

Species
Mouse
Dilution info
1/60
Notes

-

Species
Rat
Dilution info
1/60
Notes

-

Tested
Tested

Species
Rat
Dilution info
1/1000
Notes

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

Species
Human
Dilution info
1/1000
Notes

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

Expected
Expected

Species
Mouse
Dilution info
1/1000
Notes

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

Tested
Tested

Species
Human
Dilution info
1/20
Notes

For unpurified use at 1/50. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Mouse
Dilution info
1/20
Notes

For unpurified use at 1/50. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species
Rat
Dilution info
1/20
Notes

For unpurified use at 1/50. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

Select an associated product type

4 products for Alternative Product

Target data

Function

Serine kinase that plays an essential role in the NF-kappa-B signaling pathway which is activated by multiple stimuli such as inflammatory cytokines, bacterial or viral products, DNA damages or other cellular stresses (PubMed:18626576, PubMed:9244310, PubMed:9252186, PubMed:9346484). Acts as a part of the canonical IKK complex in the conventional pathway of NF-kappa-B activation and phosphorylates inhibitors of NF-kappa-B on serine residues (PubMed:18626576, PubMed:35952808, PubMed:9244310, PubMed:9252186, PubMed:9346484). These modifications allow polyubiquitination of the inhibitors and subsequent degradation by the proteasome (PubMed:18626576, PubMed:9244310, PubMed:9252186, PubMed:9346484). In turn, free NF-kappa-B is translocated into the nucleus and activates the transcription of hundreds of genes involved in immune response, growth control, or protection against apoptosis (PubMed:18626576, PubMed:9244310, PubMed:9252186, PubMed:9346484). Negatively regulates the pathway by phosphorylating the scaffold protein TAXBP1 and thus promoting the assembly of the A20/TNFAIP3 ubiquitin-editing complex (composed of A20/TNFAIP3, TAX1BP1, and the E3 ligases ITCH and RNF11) (PubMed:21765415). Therefore, CHUK plays a key role in the negative feedback of NF-kappa-B canonical signaling to limit inflammatory gene activation. 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 (PubMed:20501937). In turn, these complexes regulate genes encoding molecules involved in B-cell survival and lymphoid organogenesis. Participates also in the negative feedback of the non-canonical NF-kappa-B signaling pathway by phosphorylating and destabilizing MAP3K14/NIK. Within the nucleus, phosphorylates CREBBP and consequently increases both its transcriptional and histone acetyltransferase activities (PubMed:17434128). Modulates chromatin accessibility at NF-kappa-B-responsive promoters by phosphorylating histones H3 at 'Ser-10' that are subsequently acetylated at 'Lys-14' by CREBBP (PubMed:12789342). Additionally, phosphorylates the CREBBP-interacting protein NCOA3. Also phosphorylates FOXO3 and may regulate this pro-apoptotic transcription factor (PubMed:15084260). Phosphorylates RIPK1 at 'Ser-25' which represses its kinase activity and consequently prevents TNF-mediated RIPK1-dependent cell death (By similarity). Phosphorylates AMBRA1 following mitophagy induction, promoting AMBRA1 interaction with ATG8 family proteins and its mitophagic activity (PubMed:30217973).

Alternative names

IKKA, TCF16, CHUK, Inhibitor of nuclear factor kappa-B kinase subunit alpha, I-kappa-B kinase alpha, IKK-A, IKK-alpha, IkBKA, IkappaB kinase, Conserved helix-loop-helix ubiquitous kinase, I-kappa-B kinase 1, Nuclear factor NF-kappa-B inhibitor kinase alpha, Transcription factor 16, IKK-1, IKK1, NFKBIKA, TCF-16

Recommended products

Rabbit Recombinant Monoclonal IKK alpha antibody. Suitable for IHC-P, IP, WB, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 75 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
Y463
Purification technique
Affinity purification Protein A
Specificity

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

Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

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.

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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Immunoprecipitation - Anti-IKK alpha antibody [Y463] (ab32041), expandable thumbnail

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-IKK alpha antibody [Y463] - BSA and Azide free ab169743 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (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

    All lanes: Immunoprecipitation - Anti-IKK alpha antibody [Y463] (ab32041)

    Predicted band size: 84 kDa

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

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

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

    Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 15 µg

    Lane 2: Mouse kidney lysate at 15 µg

    Lane 3: C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

    Lane 4: Rat kidney lysate at 15 µg

    Secondary

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

    Predicted band size: 84 kDa

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

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

    All lanes: Western blot - Anti-IKK alpha antibody [Y463] (ab32041) at 1/10000 dilution

    All lanes: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate

    Secondary

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

    Predicted band size: 84 kDa

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

    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, Anti-beta Actin antibody [mAbcam 8226] - 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 Anti-beta Actin antibody [mAbcam 8226] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-IKK alpha antibody [Y463] (ab32041)

    Predicted band size: 84 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha antibody [Y463] (ab32041), expandable thumbnail

    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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (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), expandable thumbnail

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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), expandable thumbnail

    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, Goat Anti-Rabbit IgG H&L (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), expandable thumbnail

    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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (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.

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

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

    All lanes: Western blot - Anti-IKK alpha antibody [Y463] (ab32041) at 1/10000 dilution

    All lanes: Western blot - Recombinant human IKK alpha protein (Active) (Recombinant human IKK alpha protein (Active) ab102103) at 0.01 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 84 kDa

    Exposure time: 10s

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Product protocols

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