Recombinant Anti-IKK beta antibody [EPR6043] (ab124957)
![Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-256177-anti-ikk-beta-antibody-epr6043-western-blot.jpg "Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6043] to IKK beta
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-IKK beta antibody [EPR6043]
See all IKK beta primary antibodies -
Description
Rabbit monoclonal [EPR6043] to IKK beta -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
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Positive control
- HeLa and 293T cell lysates; Human cervix carcinoma and Mouse kidney tissues; HeLa cells.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 1.28 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6043 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124957 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 |
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 87 kDa).
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
(Heat to 98°C, allow to cool for 10-20 minutes) |
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| ICC/IF |
1/50 - 1/100.
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| Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
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WB
1/1000 - 1/10000. Detects a band of approximately 85 kDa (predicted molecular weight: 87 kDa). |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes) |
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ICC/IF
1/50 - 1/100. |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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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. Also phosphorylates NCOA3. -
Tissue specificity
Highly expressed in heart, placenta, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis and peripheral blood. -
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
modificationsUpon cytokine stimulation, phosphorylated on Ser-177 and Ser-181 by MEKK1 and/or MAP3K14/NIK; which enhances activity. Once activated, autophosphorylates on the C-terminal serine cluster; which decreases activity and prevents prolonged activation of the inflammatory response.
Acetylation of Thr-180 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the I-kappa-B pathway.
Ubiquitinated. Monoubiquitination involves TRIM21 that leads to inhibition of Tax-induced NF-kappa-B signaling. According to PubMed:19675099, 'Ser-163' does not serve as a monoubiquitination site. According to PubMed:16267042, ubiquitination on 'Ser-163' modulates phosphorylation on C-terminal serine residues. Monoubiquitination by TRIM21 is dirupted by Yersinia yopJ. -
Cellular localization
Cytoplasm. Membrane raft. Colocalized with DPP4 in membrane rafts. - Information by UniProt
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Database links
- Entrez Gene: 3551 Human
- Entrez Gene: 16150 Mouse
- Entrez Gene: 84351 Rat
- Omim: 603258 Human
- SwissProt: O14920 Human
- SwissProt: O88351 Mouse
- SwissProt: Q9QY78 Rat
- Unigene: 597664 Human
see all -
Alternative names
- I kappa B kinase 2 antibody
- I kappa B kinase beta antibody
- I-kappa-B kinase 2 antibody
see all
Images
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Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IKK beta knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Merged signal (red and green). Green - ab124957 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab124957 was shown to specifically react with IKK beta when IKK beta knockout samples were used. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. ab124957 and ab8245 (loading control to GAPDH) were diluted 1/500 and 1/2000 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 h at room temperature before imaging. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-139022-anti-ikk-beta-antibody-epr6043-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] (ab124957)ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Mouse kidney tissue, by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-449144-anti-ikk-beta-antibody-epr6043-western-blot-hela-edited-lysates.jpg)
Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)All lanes : Anti-IKK beta antibody [EPR6043] (ab124957) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : IKBKB CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab124957 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab124957 was shown to react with IKK beta in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264847 (CRISPR/Cas9 edited cell lysate ab257228) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and IKBKB CRISPR/Cas9 edited HeLa 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. ab124957 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at a 1 in 500 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.
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![Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-139024-anti-ikk-beta-antibody-epr6043-western-blot.jpg)
Western blot - Anti-IKK beta antibody [EPR6043] (ab124957)All lanes : Anti-IKK beta antibody [EPR6043] (ab124957) at 1/1000 dilution
Lane 1 : HeLa cell lysates
Lane 2 : 293T cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 87 kDa -
![Flow Cytometry (Intracellular) - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-270663-anti-ikk-beta-antibody-epr6043-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-IKK beta antibody [EPR6043] (ab124957)Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IKK beta with unpurified ab124957 at 1/20 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) were used as the unlabeled control.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-139023-anti-ikk-beta-antibody-epr6043-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK beta antibody [EPR6043] (ab124957)ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Human cervix carcinoma tissue, by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-139021-anti-ikk-beta-antibody-epr6043-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-IKK beta antibody [EPR6043] (ab124957)ab124957, at 1/50 dilution, staining IKK beta in HeLa cells by Immunofluorescence. -
![OI-RD Scanning - Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-139020-anti-ikk-beta-antibody-epr6043.jpg)
OI-RD Scanning - Anti-IKK beta antibody [EPR6043] (ab124957) -
![Anti-IKK beta antibody [EPR6043] (ab124957)](https://www.abcam.com/ps/products/124/ab124957/Images/ab124957-8-benefits-of-recombinant-antibodies.png)
Anti-IKK beta antibody [EPR6043] (ab124957)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (41)
ab124957 has been referenced in 41 publications.
- Cao Y et al. Decreased miR-214-3p activates NF-?B pathway and aggravates osteoarthritis progression. EBioMedicine 65:103283 (2021). PubMed: 33714889
- Gan YR et al. Dickkopf-1/cysteine-rich angiogenic inducer 61 axis mediates palmitic acid-induced inflammation and apoptosis of vascular endothelial cells. Mol Med Rep 23:N/A (2021). PubMed: 33300071
- 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
- Zhao L et al. TNF-a promotes insulin resistance in obstructive sleep apnea-hypopnea syndrome. Exp Ther Med 21:568 (2021). PubMed: 33850540
- Fang H et al. MiR-132-3p Modulates MEKK3-Dependent NF-?B and p38/JNK Signaling Pathways to Alleviate Spinal Cord Ischemia-Reperfusion Injury by Hindering M1 Polarization of Macrophages. Front Cell Dev Biol 9:570451 (2021). PubMed: 33644040