# Centrifugation speed for adherent cell line - (Jan/14/2008 )

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I am culturing mouse skin melanoma adherent cells. What is the suitable speed of centrifugation to settle down the cells? Can I use up to 1500 rpm for 5 min?
The pellet I get is dark color, not whitish pellet....is it due to this cell line is a highly pigmented cells that contain melanin pigment?

-sasoriza-

What happens if you use less rpm?
I would recommend you to centrifuge as slow as possible.

We use up to 430xg

-vista-

For most cells, we use between 100x g to 500xg for 5 min. This is sufficient to get them down.

-scolix-

QUOTE (vista @ Jan 14 2008, 04:55 AM)
What happens if you use less rpm?
I would recommend you to centrifuge as slow as possible.

We use up to 430xg

I use 1000rpm for 5 min, then when I slowly pour out the supernatant , the cell pellet is like not settle down completely..can see some still floating. I use 15 ml Falcon tubes for centrifugation.

-sasoriza-

QUOTE (scolix @ Jan 14 2008, 06:29 AM)
For most cells, we use between 100x g to 500xg for 5 min. This is sufficient to get them down.

100xg - 500xg is how many in rpm?

-sasoriza-

I see on this topic some persons need to learn how a centrifuge works and what is the difference between rpm and g !!

-Kooothor-

QUOTE (Kooothor @ Jan 14 2008, 07:06 AM)
I see on this topic some persons need to learn how a centrifuge works and what is the difference between rpm and g !!

I know there is a formula to convert g to rpm:

RCF (g) = 0.0000118 x r x rpm^2

r = radius of rotor in cm

-sasoriza-

Then why don't you give the g-force? That's the only relevant point as we don't know what centrifuge you have.

-vairus-

QUOTE (vairus @ Jan 14 2008, 08:07 AM)
Then why don't you give the g-force? That's the only relevant point as we don't know what centrifuge you have.

I need to check back what is the rotor radius of the centrifuge in my lab. I forgot how much is it. I think should be stated in the manual book. Anyway..Thanks for ur reply.

-sasoriza-

QUOTE (sasoriza @ Jan 14 2008, 10:12 AM)
I need to check back what is the rotor radius of the centrifuge in my lab. I forgot how much is it. I think should be stated in the manual book. Anyway..Thanks for ur reply.

you can simply take a ruler and measure the radius.

-scolix-

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