Recombinant Anti-IKB alpha (phospho S36) antibody [EPR6235(2)] (ab133462)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6235(2)] to IKB alpha (phospho S36)
- Suitable for: WB, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-IKB alpha (phospho S36) antibody [EPR6235(2)]
See all IKB alpha primary antibodies -
Description
Rabbit monoclonal [EPR6235(2)] to IKB alpha (phospho S36) -
Host species
Rabbit -
Specificity
This antibody detects IKB alpha only when phosphorylated at Serine 36. Stimulation may be required to allow detection of the phosphorylated protein due to low levels of endogenous expression. Please see images below for recommended treatment conditions and positive controls.
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Tested applications
Suitable for: WB, Flow Cyt (Intra)more 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
- HeLa cells treated with FBS and Calyculin A.
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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. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6235(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133462 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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WB | (1) |
1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 35 kDa).
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Flow Cyt (Intra) |
1/100.
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Notes |
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WB
1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 35 kDa). |
Flow Cyt (Intra)
1/100. |
Target
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Function
Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription. -
Involvement in disease
Ectodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant -
Sequence similarities
Belongs to the NF-kappa-B inhibitor family.
Contains 5 ANK repeats. -
Post-translational
modificationsPhosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.
Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.
Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.
Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation. -
Cellular localization
Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export. - Information by UniProt
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Database links
- Entrez Gene: 4792 Human
- Entrez Gene: 18035 Mouse
- Entrez Gene: 25493 Rat
- Omim: 164008 Human
- SwissProt: P25963 Human
- SwissProt: Q9Z1E3 Mouse
- SwissProt: Q63746 Rat
- Unigene: 81328 Human
see all -
Alternative names
- I kappa B alpha antibody
- I-kappa-B-alpha antibody
- IkappaBalpha antibody
see all
Images
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All lanes : Anti-IKB alpha (phospho S36) antibody [EPR6235(2)] (ab133462) at 1/1000 dilution (Purified)
Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours then treated with 100nM Calyculin A for 30 minutes
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) starved for 24 hours then treated with 100nM Calyculin A for 30 minutes, then the menbrane treated with Alkaline Phosphatase for 1 hour
Lane 4 : Untreated C6 (Rat glial tumor glial cell)
Lane 5 : C6 (Rat glial tumor glial cell) treated with 100ng/ml Calyculin A for 60 minutes
Lane 6 : C6 (Rat glial tumor glial cell) treated with 100ng/ml Calyculin A for 60 minutes, then the membrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
All lanes : Anti-IKB alpha (phospho S36) antibody [EPR6235(2)] (ab133462) at 1/1000 dilution (Purified)
Lane 1 : Untreated THP-1 (Human monocytic leukemia monocyte)
Lane 2 : THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml IFN-gamma for 24 hours, then 300ng/ml Brefeldin A added and treated for another 20 hours
Lane 3 : THP-1 (Human monocytic leukemia monocyte) treated with 100ng/ml IFN-gamma for 24 hours, then 300ng/ml Brefeldin A added and treated for another 20 hours, then the menbrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
Intracellular flow cytometric analysis of 4% Paraformaldehyde fixed 90% Methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) treated with 1% FBS for 3 hours then 100 nM Calyculin A for 10 minutes, labelling IKB alpha (phospho S36) with ab133462 at 1/100 dilution (1µg/ml) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody. Untreated control: Green
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (196)
ab133462 has been referenced in 196 publications.
- Li D et al. PFKFB4 promotes angiogenesis via IL-6/STAT5A/P-STAT5 signaling in breast cancer. J Cancer 13:212-224 (2022). PubMed: 34976184
- Lu F et al. Downregulation of cathepsin C alleviates endothelial cell dysfunction by suppressing p38 MAPK/NF-κB pathway in preeclampsia. Bioengineered 13:3019-3028 (2022). PubMed: 35037834
- Xie Z et al. Calcium dobesilate alleviates renal dysfunction and inflammation by targeting nuclear factor kappa B (NF-κB) signaling in sepsis-associated acute kidney injury. Bioengineered 13:2816-2826 (2022). PubMed: 35038964
- Kim YK et al. Cytokine-Induced JAK2-STAT3 Activates Tissue Regeneration under Systemic or Local Inflammation. Int J Mol Sci 23:N/A (2022). PubMed: 35216377
- Tian Y et al. Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway. Front Immunol 13:839796 (2022). PubMed: 35237277