Recombinant Anti-IKB beta antibody [EPR5037] (ab109509)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5037] to IKB beta
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-IKB beta antibody [EPR5037]
See all IKB beta primary antibodies -
Description
Rabbit monoclonal [EPR5037] to IKB beta -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, HeLa, THP-1, and MCF7 cell lysates. IP: HeLa cells. IHC-P: Human kidney tissue. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
EPR5037 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109509 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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ICC/IF |
1/50.
For unpurified use at 1/100 - 1/250. |
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Flow Cyt (Intra) |
1/20.
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 48 kDa (predicted molecular weight: 37 kDa).
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IP |
1/10 - 1/100.
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IHC-P |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
Notes |
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ICC/IF
1/50. For unpurified use at 1/100 - 1/250. |
Flow Cyt (Intra)
1/20. |
WB
1/1000 - 1/10000. Detects a band of approximately 48 kDa (predicted molecular weight: 37 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250. |
Target
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Function
Inhibits NF-kappa-B by complexing with and trapping it in the cytoplasm. However, the unphosphorylated form resynthesized after cell stimulation is able to bind NF-kappa-B allowing its transport to the nucleus and protecting it to further NFKBIA-dependent inactivation. Association with inhibitor kappa B-interacting NKIRAS1 and NKIRAS2 prevent its phosphorylation rendering it more resistant to degradation, explaining its slower degradation. -
Tissue specificity
Expressed in all tissues examined. -
Sequence similarities
Belongs to the NF-kappa-B inhibitor family.
Contains 6 ANK repeats. -
Post-translational
modificationsPhosphorylated by RPS6KA1; followed by degradation. Interaction with NKIRAS1 and NKIRAS2 probably prevents phosphorylation. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4793 Human
- Omim: 604495 Human
- SwissProt: Q15653 Human
- Unigene: 9731 Human
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Alternative names
- I kappa B beta antibody
- I-kappa-B-beta antibody
- IkappaBbeta antibody
see all
Images
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Anti-IKB beta antibody [EPR5037] (ab109509) at 1/1000 dilution (Purified) + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted? -
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: IKB beta knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - unpurified ab109509 observed at 45 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab109509 was shown to specifically react with IKB beta in wild type cells as signal was lost in IKB beta knockout cells. Wild-type and IKB beta knockout samples were subjected to SDS-PAGE. ab109509 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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ab109509 (purified) at 1:20 dilution (1 µg) immunoprecipitating IKB beta in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab109509 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109509 in HeLa whole cell lysateFor western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
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Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling IKB beta with purified ab109509 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). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling IKB beta with purified ab109509 at 1:50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB beta antibody [EPR5037] (ab109509)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling IKB beta with purified ab109509 at 1:500 dilution (0.20 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB beta antibody [EPR5037] (ab109509)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue using unpurified ab109509 at a dilution of 1/100.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-IKB beta antibody [EPR5037] (ab109509) at 1/1000 dilution (Unpurified)
Lane 1 : Jurkat cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : THP-1 cell lysate
Lane 4 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 37 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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Datasheet download
References (7)
ab109509 has been referenced in 7 publications.
- Shen L et al. A naturally derived small molecule NDSM253 inhibits IKK1 to suppress inflammation response and promote bone healing after fracture. Am J Transl Res 13:24-37 (2021). PubMed: 33527006
- Xiao N et al. Egg yolk oils exert anti-inflammatory effect via regulating Nrf2/NF-?B pathway. J Ethnopharmacol 274:114070 (2021). PubMed: 33785356
- Zhao L et al. TNF-a promotes insulin resistance in obstructive sleep apnea-hypopnea syndrome. Exp Ther Med 21:568 (2021). PubMed: 33850540
- Liu J et al. Long noncoding RNA LINC01578 drives colon cancer metastasis through a positive feedback loop with the NF-?B/YY1 axis. Mol Oncol 14:3211-3233 (2020). PubMed: 33040438
- Hsueh YS et al. Nuclear KIT induces a NFKBIB-RELA-KIT autoregulatory loop in imatinib-resistant gastrointestinal stromal tumors. Oncogene 38:6550-6565 (2019). PubMed: 31363162
- Wen H et al. Recurrent ECSIT mutation encoding V140A triggers hyperinflammation and promotes hemophagocytic syndrome in extranodal NK/T cell lymphoma. Nat Med 24:154-164 (2018). WB . PubMed: 29291352
- Yi L et al. Inflammation-mediated SOD-2 upregulation contributes to epithelial-mesenchymal transition and migration of tumor cells in aflatoxin G1-induced lung adenocarcinoma. Sci Rep 7:7953 (2017). PubMed: 28801561