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APPROVED PMLSC ASSAY PROTOCOLS
  • PMLSC Assay Protocol Review Document for MKP-1 HTS
  • PMLSC Assay Protocol Review Document for Cdc25B Catalytic Domain HTS
  • GNHR Translocation Assay APAC Review document
  • Necroptosis PMLSC APAC Document
  • PLK-1 PBD PMLSC APAC Document
  • West Nile Virus NS2bNS3 PMLSC APAC Document
  • Time Resolved Fluorescence Resonance Energy Transfer assay for Plk1 inhibitors
  • PKD IMAP TR-FRET APAC Document
  • High-content cell-based screening for modulators of autophagy
  • MKP-3 Chemical Complementation assay
  • PMLSC PROBE REPORTS
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    Approved PMLSC Assay Protocols
     
    bullet point  PMLSC Assay Protocol Review Document for MKP-1 HTS
     
     
    The mitogen-activated protein kinases (MAPK) are members of the signaling cascades for diverse extracellular stimuli that regulate fundamental cellular processes including embryogenesis, differentiation, mitosis, apoptosis, movement and gene expression. Four distinct MAP kinase families have been described; the extracellular signal-regulated kinases (ERK’s), c-jun N-terminal (JNK) or stress-activated protein kinases (SAPK), ERK5/big MAP kinase 1 (BMK1), and the p38 group of protein kinases. Since MAPK’s are regulated by phosphorylation, it therefore follows that phosphatases will be a key element of their control, and a family of dual specificity (Ser/Thr/Tyr) MAPK phosphatases (MKP1, MKP2 and MKP3) have been implicated. Mitogen-activated protein kinase phosphatase-1 (MKP-1, DUSP1 or CL100) dephosphorylates and inactivates MAPK substrates, such as p38, JNK, and Erk, and has been implicated in neoplasia. The MKP-1 assay is a homogeneous fluorescence intensity assay measuring the hydrolysis of the generic substrate 3-O-methylfluorescein phosphate (OMFP). We report here the optimization that was required for the development of a 384-well HTS assay that conforms to the PMLSC assay development and implementation guidelines.
     
    Documents
  • PMLSC Assay Protocol Review Document for MKP-1 HTS
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