1 reply to this topic

[Dark Monarch]

I've immortalized hepatocytes from neonate Black 6 mice.  I'm culturing them in DMEM plus 10%FCS and hormones(insulin, dexamethasone, EGF). To show that they are indeed hepatocytes, I'm looking at albumin synthesis.  I find albumin in the medium but when I lyse the cells with RIPA buffer, I only find a band at ~120,000kDa, not at 66,000kDa where the albumin migrates.
Is it the medium; is it the RIPA lysing buffer?
PS They don't look like your normal hepatocytes.

[bob1]

Occam's razor says that the simplest answer is usually the correct one... Perhaps they aren't hepatocytes.  I would do some morphology and histochemistry for cell specific markers to make sure that they are hepatocytes.