PhosphoTracer IKKα pSer176/180 + Total ELISA Kit (ab119662)
- Product namePhosphoTracer IKKα pSer176/180 + Total ELISA Kit
- Tests1 x 96 well plate
- Sample typeCell culture extracts
- Assay typeSandwich
- Assay time1h 15m
- Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat
- Product overview
PhosphoTracer assays use a traditional immuno-sandwich format, but with a major difference both the analyte and the assay reagents are added to the PhosphoTracer assay microplate at the same time. After a short incubation period, unbound assay reagents and analytes are washed away, and immuno-complexes containing both antibodies are detected. The process can take as little as 60 minutes to complete.
PhosphoTracer kits also allow a higher degree of assay flexibility. In contrast to other ELISA formats, no antibodies are present on the assay microplate itself, so assays for several different targets can be performed in different wells on the same microplate. Simply mix the lysate with your target reagents of choice, using the microplate configuration of your choice.
A whole new way of performing cellular assays, PhosphoTracer takes the hard work out of running a standard ELISA, while still giving the high quality results expected from a sandwich immunoassay. Fully self-contained kits are supplied in convenient 96-well packs. Simple to use and highly sensitive PhosphoTracer kits are designed to get results, fast.
Abcam's PhosphoTracer IKKalpha assay kits detect endogenous levels of IKKalpha (GenBank Accession NP_001269) in cellular lysates. Phospho-IKKalpha assay kits detect IKKalpha only when phosphorylated at Ser176/180. Total IKKalpha assay kits detect IKKalpha irrespective of phosphorylation status.
The substrate used with the HRP conjugated detection antibody is a combination of 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) (wavelength exc/em = 530-540nm / 590-600nm), a highly sensitive and stable substrate for HRP) and ADHP Dilution Buffer (a stabilized H2O2 solution). Learn more about the fluorogenic substrate, ADHP.
Sensitivity: Phospho-IKKalpha: 0.25ng/ml (tested with rhIKKalpha), IKKalpha: 0.5ng/ml (tested with rhIKKalpha).
Range: Phospho-IKKalpha: 0.25 - 250 ng/ml (tested with rhIKKalpha), IKKalpha: 0.5 - 250 ng/ml (tested with rhIKKalpha).
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 96-well PhosphoTracer assay plate (stripwell) 1 unit Adherent plate seal 2 units ADHP (100X) 1 x 120µl ADHP Dilution Buffer 1 x 15ml Assay Control Lysate (lyophilized) 1 x 0.25ml Enhancer Solution 1 x 1ml Lysis Buffer (5X) 1 x 15ml Mouse monoclonal IKKalpha (HRP) (24 assay points) 1 x 0.75ml Mouse monoclonal Phospho-IKKalpha (HRP) (72 assay points) 3 x 0.75ml Rabbit monoclonal IKKalpha (24 assay points) 1 x 0.75ml Rabbit monoclonal Phospho-IKKalpha (Ser176/180) (72 assay points) 3 x 0.75ml Stop Solution 1 x 2ml Wash Buffer (10X) 1 x 15ml
- Research Areas
Contains 1 protein kinase domain.
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.
- chukConserved helix-loop-helix ubiquitous kinaseI-kappa-B kinase 1
- I-kappa-B kinase alphaIkappaB kinaseIkB kinase alpha subunitIkBKAIKK a kinaseIKK-AIKK-alphaIKK1IKKAIKKA_HUMANInhibitor of nuclear factor kappa-B kinase subunit alphaNFKBIKANuclear factor NF-kappa-B inhibitor kinase alphaNuclear factor NFkappaB inhibitor kinase alphaTCF-16TCF16Transcription factor 16
Our Abpromise guarantee covers the use of ab119662 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||sELISA. Tested in HeLa.|
PhosphoTracer IKKα pSer176/180 + Total ELISA Kit images
Using the IKKalpha assay kits, a significant stimulation of IKKalpha phosphorylation at Ser176/180 is detected in HeLa cells treated with TNF alpha for 5 minutes compared with untreated cells, while no change in total IKKalpha levels is observed over this time period.
A dilution series of recombinant active IKKalpha was prepared by diluting to various concentrations in Lysis Mix, containing 0.1% BSA. These dilution series were analyzed using the phospho-IKKalpha assay. Limit of detection of active rhIKKalpha was around 250 pg/mL. No cross-reactivity with active rhIKKB was detected over this concentration range.
References for PhosphoTracer IKKα pSer176/180 + Total ELISA Kit (ab119662)
ab119662 has not yet been referenced specifically in any publications.