Recombinant Anti-IKK alpha antibody [Y463] - BSA and Azide free (ab169743)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y463] to IKK alpha - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-IKK alpha antibody [Y463] - BSA and Azide free
See all IKK alpha primary antibodies -
Description
Rabbit monoclonal [Y463] to IKK alpha - BSA and Azide free -
Host species
Rabbit -
Specificity
The rat recommendation is based on the WB results. We do not guarantee IHC-P for rat.
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Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1-WT; Daudi,RAW 264.7 and C6 whole cell lysate. Mouse and rat kidney lysate. IP: HeLa cell lysate; IHC-P: Human ovarian cancer tissue and Mouse kidney tissue; Flow Cyt (intra): HAP1 wildtype and Daudi cells.
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General notes
ab169743 is the carrier-free version of ab32041.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 +4°C. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y463 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab169743 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 88 kDa (predicted molecular weight: 85 kDa).
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 88 kDa (predicted molecular weight: 85 kDa). |
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. 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. -
Tissue specificity
Widely expressed. -
Involvement in disease
Defects 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. -
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
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 localization
Cytoplasm. 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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All lanes : Anti-IKK alpha antibody [Y463] (ab32041) at 1/1000 dilution (Purified)
Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : Mouse kidney lysate
Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysate
Lane 4 : Rat kidney lysate
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 85 kDa -
Anti-IKK alpha antibody [Y463] (ab32041) at 1/10000 dilution + Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution
Predicted band size: 85 kDa -
This data was developed using ab32041, the same antibody clone in a different buffer formulation.
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 ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. 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. -
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 (ab172730) instead of ab169743 in HeLa whole cell lysate.
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
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32041).
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This data was developed using ab32041, the same antibody clone in a different buffer formulation. 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, 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).
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This data was developed using ab32041, the same antibody clone in a different buffer formulation.
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 ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. 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.
Protocols
Datasheets and documents
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Datasheet download
References (3)
ab169743 has been referenced in 3 publications.
- Kunishige R et al. Targeted protein degradation by Trim-Away using cell resealing coupled with microscopic image-based quantitative analysis. Front Cell Dev Biol 10:1027043 (2022). PubMed: 36601537
- Kiss L et al. A tri-ionic anchor mechanism drives Ube2N-specific recruitment and K63-chain ubiquitination in TRIM ligases. Nat Commun 10:4502 (2019). PubMed: 31582740
- Clift D et al. A Method for the Acute and Rapid Degradation of Endogenous Proteins. Cell 171:1692-1706.e18 (2017). PubMed: 29153837