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In many cell lines, MMP-9 is expressed at low levels. However, treatment with phorbol myristate acetate (PMA) – a protein kinase C ligand – markedly stimulates MMP9 expression (Roomi et al., 2009). For analysis of MMP-9 in cell lines, it may be necessary to first stimulate your cells with PMA to detect a signal.
The following protocol has been optimized for use with U937 cells.
Lysate can either be stored at -80°C or used immediately for western blotting.
Add proteinase inhibitor before use.
|NP-40 or Triton X-100||1.0%|
|SDS (sodium dodecyl sulfate)||0.1%|
|Tris, pH 8.0||50 mM|