Hello everyone,
My experiments require me to form a 0.5 wt% agarose with a certain chemokine on a 24 well plate. This would allow slower release of the chemokine and we want to check the migration of cells from the upper side of a transwell insert to the lower part due to the chemokine released from agarose.
The agarose gel seems to float when media/PBS is added. Is there a simple way to prevent it.
I wouldn´t like to coat the well plate as it may effect the cell migration study.
Thanks in advance.
Laurence Stuart Dawkins-Hall
- When I used to coat transwells for similar reasons, I would add about 100ul of 1% gelatin and incubate for 15min @ 37C before removing excess and then incubating for a further 15min
- You could try gelatin instead of agarose - they are used for the same reason - but I suspect the problemn s that you are caoting with agarose but not then removing excess, culminating in a residual coat:
- For this sort of molecular barrier you only need a microfilm coating, which you will obtain by allowing to incubate and permeate poars before removing overlysng colloid with a pipette: That will leave enough to create a barrier but not so much that it will float into suspension
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