Hi all,
I want to do western blot for one of my secretome proteins.
I thought to use viva spin in order to concentrate the medium.
How should I proceed? can I add lysis buffer? how much?
or should I do pre-rinse first?
If anyone can help me it will be great and really appreciate.
Shani
Hi there,
What you have to check first is the chemical compatibility of your buffer with the membrane and then to respect the centrifugation condition (see the vivaspin leaflet for instructions).The rate of filtration will depend on the nature of the sample so the time needed to get the right protein concentration will be variable from one sample to the other.
Hi Shani,
I attached the guidelines for usage with a buffer compatibility table. Besides that, it also depends what kind of viva spin you have. Which volume do you want to concentrate, what kind of a cut off do you need. In general for Viva Spin 500, we start with an equilibriation step with water (5mins, 14000g). Then the same with your buffer. Afterwards, we carefully check different speeds (from 3000g-14000g and 3-10 mins) depending on the amount of the sample. I hope this will help you.
Best,
Can
Dear Shani:
http://www.vivaproducts.com/downloads/vivaspin-2-500-operating-instructions.pdf
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