Need Some Help Please - (May/06/2007 )
Im developing an assay that requires a mild proteinase digestion of fixed cells- problem is i need to wash away all the digested material and retain the intact cells - anyone know how i can do this? thanks so much
-BioDude1-
Hi!
Can you trypsinize cells, use proteinase then spin though a sucrose gradient?
Good Luck!
QUOTE (BioDude1 @ May 6 2007, 12:17 PM)
Im developing an assay that requires a mild proteinase digestion of fixed cells- problem is i need to wash away all the digested material and retain the intact cells - anyone know how i can do this? thanks so much
-biokmst-
QUOTE (BioDude1 @ May 6 2007, 06:17 PM)
Im developing an assay that requires a mild proteinase digestion of fixed cells- problem is i need to wash away all the digested material and retain the intact cells - anyone know how i can do this? thanks so much
does your mild protease digestion detaches cells from matrix? so, if not, the still fixed cells can be washed with medium or PBS;
if cells are detached, elutration would be nice (only usefull for larger amounts of cells); cell sorting or Nycodenz gradient centrifugarion may also be nice...
-The Bearer-