Human Phospho-PKMYT1 (T495) and Total PKMYT1 PharmaGenie ELISA Kit (SBRS1931)

The Phospho-PKMYT1 (T495) and Total PKMYT1 ELISA Kit from Assay Genie enables researchers to quantify both phosphorylated PKMYT1 at threonine 495 and total PKMYT1 protein in parallel from the same lysate, supporting normalisation against total antigen when studying mitotic entry and G2/M checkpoint signalling. PKMYT1 is a membrane-associated serine/threonine kinase that phosphorylates CDK1 on Thr14 and Tyr15, restraining CDK1-cyclin B activity until mitotic onset; autophosphorylation at T495 is widely used as a readout of PKMYT1 catalytic activity. The kit is therefore relevant to investigations of the DNA damage response, mitotic catastrophe in p53-deficient tumour cells, and screening of small-molecule PKMYT1 inhibitors as synthetic-lethal targets. The 96-assay format is supplied with pre-coated microplate wells, lyophilised standards for both analytes, biotinylated detection antibodies, streptavidin-HRP conjugate, TMB substrate, and assay diluents compatible with cell lysates and tissue lysates prepared in standard RIPA buffer. The assay has been validated for human, mouse, and rat PKMYT1 with a recommended sample volume of 50 µL per well loaded in duplicate, and standard curves are routinely prepared between 0.156 ng/mL and 10 ng/mL for both the phospho-T495 and total PKMYT1 readouts. Typical research-use workflows include serum-starved HeLa or U2OS cells released from a double-thymidine block, treatment with doxorubicin or etoposide to induce checkpoint activation, and comparison of phospho-T495 versus total PKMYT1 across treatment arms. Storage recommendations include keeping the sealed kit components at 4 °C and reserving the standard aliquots at -20 °C to preserve activity through multiple freeze-thaw cycles.