Rabbit Recombinant Monoclonal IKK beta antibody. Carrier free. Suitable for IHC-P, WB and reacts with Mouse, Human samples.
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Expected | Not recommended | Tested | Not recommended | Not recommended |
Mouse | Tested | Not recommended | Expected | Not recommended | Not recommended |
Rat | Predicted | Not recommended | Predicted | Not recommended | Not recommended |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes (Heat to 98°C, allow to cool for 10-20 minutes) Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Human, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Can be blocked with ab154148 |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info - | Notes - |
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Serine kinase that plays an essential role in the NF-kappa-B signaling pathway which is activated by multiple stimuli such as inflammatory cytokines, bacterial or viral products, DNA damages or other cellular stresses (PubMed:20434986, PubMed:20797629, PubMed:21138416, PubMed:30337470, PubMed:9346484). Acts as a part of the canonical IKK complex in the conventional pathway of NF-kappa-B activation (PubMed:9346484). Phosphorylates inhibitors of NF-kappa-B on 2 critical serine residues (PubMed:20434986, PubMed:20797629, PubMed:21138416, PubMed:9346484). These modifications allow polyubiquitination of the inhibitors and subsequent degradation by the proteasome (PubMed:20434986, PubMed:20797629, PubMed:21138416, PubMed:9346484). In turn, free NF-kappa-B is translocated into the nucleus and activates the transcription of hundreds of genes involved in immune response, growth control, or protection against apoptosis (PubMed:20434986, PubMed:20797629, PubMed:21138416, PubMed:9346484). In addition to the NF-kappa-B inhibitors, phosphorylates several other components of the signaling pathway including NEMO/IKBKG, NF-kappa-B subunits RELA and NFKB1, as well as IKK-related kinases TBK1 and IKBKE (PubMed:11297557, PubMed:14673179, PubMed:20410276, PubMed:21138416). IKK-related kinase phosphorylations may prevent the overproduction of inflammatory mediators since they exert a negative regulation on canonical IKKs (PubMed:11297557, PubMed:20410276, PubMed:21138416). Phosphorylates FOXO3, mediating the TNF-dependent inactivation of this pro-apoptotic transcription factor (PubMed:15084260). Also phosphorylates other substrates including NAA10, NCOA3, BCL10 and IRS1 (PubMed:17213322, PubMed:19716809). Phosphorylates RIPK1 at 'Ser-25' which represses its kinase activity and consequently prevents TNF-mediated RIPK1-dependent cell death (By similarity). Phosphorylates the C-terminus of IRF5, stimulating IRF5 homodimerization and translocation into the nucleus (PubMed:25326418). Following bacterial lipopolysaccharide (LPS)-induced TLR4 endocytosis, phosphorylates STAT1 at 'Thr-749' which restricts interferon signaling and anti-inflammatory responses and promotes innate inflammatory responses (PubMed:38621137). IKBKB-mediated phosphorylation of STAT1 at 'Thr-749' promotes binding of STAT1 to the ARID5A promoter, resulting in transcriptional activation of ARID5A and subsequent ARID5A-mediated stabilization of IL6 (PubMed:32209697). It also promotes binding of STAT1 to the IL12B promoter and activation of IL12B transcription (PubMed:32209697).
IKKB, IKKB, IKBKB, Inhibitor of nuclear factor kappa-B kinase subunit beta, I-kappa-B-kinase beta, IKK-B, IKK-beta, IkBKB, I-kappa-B kinase 2, Nuclear factor NF-kappa-B inhibitor kinase beta, Serine/threonine protein kinase IKBKB, IKK-2, IKK2, NFKBIKB
Rabbit Recombinant Monoclonal IKK beta antibody. Carrier free. Suitable for IHC-P, WB and reacts with Mouse, Human samples.
IgG
Rabbit
Constituents: PBS
Liquid
Monoclonal
Yes
EPR6043
Affinity purification Protein A
Blue Ice
+4°C
+4°C
Do Not Freeze
ab171364 is the carrier-free version of Anti-IKK beta antibody [EPR6043] ab124957.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
This WB data was generated using the same anti-IKK beta antibody clone [EPR6043] in a different buffer formulation (cat# 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 - Anti-IKK beta antibody [EPR6043] ab124957 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-IKK beta antibody [EPR6043] 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. Anti-IKK beta antibody [EPR6043] ab124957 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-IKK beta antibody [EPR6043] (Anti-IKK beta antibody [EPR6043] ab124957)
Predicted band size: 51 kDa, 87 kDa
Anti-IKK beta antibody [EPR6043] ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Mouse kidney tissue, by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-IKK beta antibody [EPR6043] ab124957).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation (Anti-IKK beta antibody [EPR6043] ab124957).
Lanes 1- 2: Merged signal (red and green). Green - Anti-IKK beta antibody [EPR6043] ab124957 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-IKK beta antibody [EPR6043] 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 Human IKBKB (IKK beta) knockout HeLa cell line ab264847 (CRISPR/Cas9 edited cell lysate Human IKBKB (IKK beta) knockout HeLa 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. Anti-IKK beta antibody [EPR6043] ab124957 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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.
All lanes: Western blot - Anti-IKK beta antibody [EPR6043] (Anti-IKK beta antibody [EPR6043] ab124957) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: IKBKB CRISPR/Cas9 edited HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 90 kDa
Anti-IKK beta antibody [EPR6043] ab124957, at 1/100 dilution staining IKK beta in paraffin-embedded Human cervix carcinoma tissue, by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-IKK beta antibody [EPR6043] ab124957).
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
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