Wash L929 Cells with DPBS (with or w/out CA MG?)
Posted 22 February 2010 - 05:23 PM
I am doing cytotoxicity testing using Bio-Rad DC protein Test kit where L929 mouse fibroblast cells are cultured in 24Well Microplate.
Before the testing, medium are removed and washed with DPBS (Room Temp) 3 times before incubating them with lysate (to lyse the cells)
OK, now, i have been using DPBS (withOUT CA,MG) to wash the cells, and stained the cells to look for the cells distribution layer over the well.
Recently i changed the PBS to PAA brand, and the cells Peels Off (almost half) after washing them.
This problem does not exist previously.
Should i use DPBS with CA,MG? I wont be doing trypsinization at this stage, so CA,MG present won't is not a matter , am i correct?
CA,MG will causes the protein binding more strongly, will this eventually causes the protein content to increase in my protein assay?
Or should i just stick back with my PBS w/out CA Mg, and warmed it before washing?
Posted 23 February 2010 - 10:06 AM
Posted 01 January 2012 - 01:02 AM
Edited by lamaksha77, 01 January 2012 - 01:02 AM.