p38 MAPK alpha pThr180/Tyr182 + Total PhosphoTracer ELISA Kit (ab119665)
- Product namep38 MAPK alpha pThr180/Tyr182 + Total PhosphoTracer ELISA KitSee all p38 MAPKalpha (phospho Thr180/Tyr182)+ total p38 MAPKalpha kits ...
- Detection methodFluorescent
- Tests1 x 96 well plate
- Sample typeCell culture extracts
- Assay typeSemi-quantitative
- Assay time2h 0m
- Assay durationOne step assay
- 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 p38 MAPKalpha assay kits detect endogenous levels of p38 MAPKalpha (GenBank Accession NP_001306) in cellular lysates. The phospho-p38 MAPKalpha assay detects p38 MAPKalpha only when phosphorylated at Thr180/Tyr182. The total p38 MAPKalpha assay detects p38 MAPKalpha 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-p38 MAPKalpha: 0.1 ng/ml (tested with rh p38 MAPK), p38 MAPKalpha: 1 ng/ml (tested with rh p38 MAPK).
Range: Phospho-p38 MAPKalpha: 0.1-250 ng/ml (tested with rh p38 MAPK), p38 MAPKalpha: 1-250 ng/ml (tested with rh p38 MAPK).
- 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 p38 MAPKalpha (24 assay points) 1 x 0.75ml Mouse monoclonal Phospho-p38 MAPKalpha (Thr180/Tyr182) (72 assay points) 3 x 0.75ml Rabbit polyclonal p38 MAPKalpha (HRP) (24 assay points) 1 x 0.75ml Rabbit polyclonal Phospho-p38 MAPKalpha (HRP) (72 assay points) 3 x 0.75ml Stop Solution 1 x 2ml Wash Buffer (10X) 1 x 15ml
- FunctionResponds to activation by environmental stress, pro-inflammatory cytokines and lipopolysaccharide (LPS) by phosphorylating a number of transcription factors, such as ELK1 and ATF2 and several downstream kinases, such as MAPKAPK2 and MAPKAPK5. Plays a critical role in the production of some cytokines, for example IL-6. May play a role in stabilization of EPO mRNA during hypoxic stress. Isoform Mxi2 activation is stimulated by mitogens and oxidative stress and only poorly phosphorylates ELK1 and ATF2. Isoform Exip may play a role in the early onset of apoptosis.
- Tissue specificityBrain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.
- Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain.
- DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
modificationsDually phosphorylated on Thr-180 and Tyr-182, which activates the enzyme.
Phosphorylated upon DNA damage, probably by ATM or ATR.
- Cellular localizationCytoplasm. Nucleus.
- CSAID binding protein
- CSAID-binding protein
- Csaids binding protein
- Cytokine suppressive anti-inflammatory drug-binding protein
- MAP kinase 14
- MAP kinase MXI2
- MAP kinase p38 alpha
- MAPK 14
- MAX-interacting protein 2
- Mitogen-activated protein kinase 14
- Mitogen-activated protein kinase p38 alpha
- p38 MAP kinase
- p38 mitogen activated protein kinase
- p38alpha Exip
- Stress activated protein kinase 2A
Our Abpromise guarantee covers the use of ab119665 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, THP1, Jurkat.|
p38 MAPK alpha pThr180/Tyr182 + Total PhosphoTracer ELISA Kit images
As shown, using the p38 MAPKalpha assay kits, a significant stimulation of p38 MAPKalpha phosphorylation at Thr180/Tyr182 is detected in HEK293 cells treated with anisomycin for 30 minutes compared with untreated cells, while no change in total p38 MAPKalpha levels is observed.
Using the PhosphoTracer phospho-p38 MAPKalpha assay, the length of assay incubation time that was required to detect recombinant phospho-p38 MAPKalpha was examined. After 15 mins, phospho-p38 MAPKalpha concentrations of approximately 4 ng/mL were readily detected, while after 1 hour incubation, the limit of detection was approximately 1 ng/mL, and with a much improved assay window. Incubation times longer than one hour showed a slight improvement on assay performance.
PhosphoTracer p38 MAPKalpha (pT180/Y182) + total p38 MAPKalpha ELISA Kit (ab119665) used in Sandwich ELISA.
References for p38 MAPK alpha pThr180/Tyr182 + Total PhosphoTracer ELISA Kit (ab119665)
ab119665 has not yet been referenced specifically in any publications.