Assay phosphorylated MEK-1 ([pSer218/Ser222] MAPK-ERK kinase 1, pMEK1) levels in cell lysates of human, mouse, and rat origin with our Thermo Scientific Phospho MEK1 ELISA Kit.
This phospho-MEK1 ELISA kit is an immunoassay for the quantitative determination of phosphorylated MEK-1 ([pSer218/Ser222] MAPK-ERK kinase 1, pMEK1) in cell lysates. A recombinant MEK1 standard is provided with the kit. The assay uses a monoclonal antibody to MEK1 precoated in a microplate to bind the pMEK in the standards or sample. After a short incubation, the excess sample or standard is washed out of the plate and a phospho-MEK-1/2 detection antibody is added. This antibody binds to the pMEK captured on the plate. After a short incubation, the excess antibody is washed out and secondary conjugate is added, which binds to the phospho-MEK detection antibody. Excess conjugate is washed out and TMB substrate is added. After a short incubation, the enzyme reaction is stopped and the absorbance is read at 450nm. The measured optical density is directly proportional to the concentration of pMEK in either standards or samples.
- Target: phosphorylated MEK-1 ([pSer218/Ser222] MAPK-ERK kinase 1, pMEK1, phospho-MEK1)
- Format: colorimetric ELISA (TMB substrate) in clear 96-well strip plate
- Measurement: absorbance at 450nm minus absorbance at 550nm
- Assay Range: 187.5 to 6000pg/mL
- Sensitivity: < 85.2pg/mL
- Precision: dependent upon sample type and level (see instructions)
- Sample Types: cell lysates of human, mouse, and rat origin
- Total Assay Time: 3 hours
- Sample Size: 100μL
- Specificity: no significant cross-reaction with non-phosphorylated MEK-1, phospho-JNK, ERK2 and phospho-ERK2; approx. 12% cross-reaction with phospho-MEK-2 (pMEK2).
Complete kit contains pre-coated antibody-plate, detection antibodies, buffers, standard, substrate, and stop solution.