HEK293 cells fractionation

[shinysky]

Hi. I am working on fractionation of HEK293 cells to get cytosolic, membrane and nuclear fractions.

Before fractionation, I washed HEK293 cells using PBS and spin-downed them at 100g.
When I loaded the supernatant from this PBS wash and spin-down step as a control,
a cytosolic marker (LDH) was detected, to my surprise. More surprisingly, when I
spin-downed HEK293 cells at 700g instead of 100g, a nuclear marker (HDAC2) was
detected, along with LDH.

It suggested to me that HEK293 cells were crushed during spin-down (even at 100g).

Is it normal? Or are my HEK293 cells somehow wrong? I could not find any related
information on the web, which is why I am writing here.

Thanks in advance.

[harrygail87]

hi
hello everyone i am new . i want to say about HEK293 cells fractionation. The cell polarity regulator, human Scribble (hScrib), is a potential tumour suppressor whose loss is a frequent event in late-stage cancer development. hese results provide a clear mechanistic explanation of how hScrib can regulate ERK signalling and begin to explain how loss of hScrib during cancer development can contribute to disease progression.