Anti-IKK alpha (phospho T23) antibody (ab38515)
- Datasheet
- References (13)
- Protocols
Overview
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Product nameAnti-IKK alpha (phospho T23) antibody
See all IKK alpha primary antibodies -
DescriptionRabbit polyclonal to IKK alpha (phospho T23)
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Host speciesRabbit
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SpecificityThis antibody is specific for IKK alpha only when phosphorylated at threonine 23.
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Tested applicationsSuitable for: ICC/IF, WB, IHC-P, ELISA, IHC-Frmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic phosphopeptide derived from Human IKK alpha around the phosphorylation site of threonine 23 (LGTpGG).
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Positive control
- Colon carcinoma tissue slides and MDA-MB-435 cell extract.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesThe antibody was affinity purified from rabbit antiserum by affinity chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab38515 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 |
|---|---|---|
| ICC/IF | Use a concentration of 1 µg/ml. | |
| WB | 1/500 - 1/1000. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa). | |
| IHC-P | Use at an assay dependent concentration. | |
| ELISA | 1/4000. | |
| IHC-Fr | Use at an assay dependent concentration. See Abreview. |
Target
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FunctionActs 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.
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Tissue specificityWidely expressed.
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Involvement in diseaseDefects 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.
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Sequence similaritiesBelongs to the protein kinase superfamily. Ser/Thr protein kinase family. I-kappa-B kinase subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated 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 localizationCytoplasm. Nucleus. Shuttles between the cytoplasm and the nucleus.
- Information by UniProt
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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
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Alternative names
- chuk antibody
- CHUK1 antibody
- Conserved Helix Loop Helix Ubiquitous Kinase antibody
see all
Images
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Western blot - Anti-IKK alpha (phospho T23) antibody (ab38515)All lanes : Anti-IKK alpha (phospho T23) antibody (ab38515) at 1/500 dilution
Lane 1 : MDA-MB-435 cell extract.
Lane 2 : MDA-MB-435 cell extract + EGF ( The cells were treated with 200ng/ml EGF for 5 minutes).
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Alkaline Phosphatase AffiniPure Goat Anti-Rabbit IgG (H+L)
Predicted band size: 85 kDa
Observed band size: 85 kDa
Lanes can be loaded with 5-30µg of total protein. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)ab38515 at a 1:50 dilution staining IKK alpha in Human colon carcinoma tissue using Immunohistochemistry, Paraffin Embedded Tissue.Left image : untreated.
Right image : treated with phosphopeptide.
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Immunohistochemistry (Frozen sections) - Anti-IKK alpha (phospho T23) antibody (ab38515)This image is courtesy of an anonymous Abreviewab38515 at 1/100 staining mouse skin tissue sections by IHC-Fr. The tissue was paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. An Alexa Fluor ® conjugated goat anti-rabbit antibody was used as the secondary.
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Immunocytochemistry/ Immunofluorescence - Anti-IKK alpha (phospho T23) antibody (ab38515)ICC/IF image of ab38515 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab38515, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
References
This product has been referenced in:
- Dong ZW & Yuan YF Juglanin suppresses fibrosis and inflammation response caused by LPS in acute lung injury. Int J Mol Med 41:3353-3365 (2018). Read more (PubMed: 29532887) »
- Zhang H et al. Neuroprotective effects of isosteviol sodium through increasing CYLD by the downregulation of miRNA-181b. Brain Res Bull 140:392-401 (2018). WB ; Mouse . Read more (PubMed: 29807144) »
