Protein is in PBS (40mM phosphate buffer and 100mM NaCl). Dialysis performed at 4 degree Celcius.
Thanks!
I would let it go overnight, making sure to keep the dialysis bag or cassette in rapid motion by stirring to keep the sample inside well-mixed.
Dear Devansu
to reach the equilibrium 8h generally are enough but is also depends from the pose size of your membrane.. big pores..fast exchange. however how much Salt do you remove depends from the ratio from inside and outside volume. for example in your case if
you perform a dialysis of 10ml in 1liter for example your ratio is 100 thatmrans at the end also if you wait 24h you still have 1mM of Salt and of you would like to go down you need to perform a second step of dilalisys ( generally 1 step 8h during the day and 2nd step over night.
alternativelly you can use desalting which is certanilly faster and in a single step can completelly remove the salts.
you can find more details about it on my blog : ProteoCool https://proteocool.blogspot.com/
ProteoCool n°8 (Buffer Exchange Methods overview)
and
ProteoCool n°11(Rapid buffer Exchange by PD-10)
available at Page3.
good luck
Manuele
Dear Devanshu Mehta
The size of the pore is quite relevant. Even though, as previously suggested, 1 step 8h during the day and 2nd step over night work pretty well for small volumes of sample (lower than 25 mL) in at least 500 mL of buffer.
Kind regards,
Rosa María
Hi Devanshu,
I usually leave the dialysis bag overnight in the stirring process and change the dialysis medium frequently.
It depend on sample volume, buffer volume ( usually i use buffer volume = 300*sample volume), dialysis mode( mixe or no), temperature, type of dialysis product (cassete or tubing, the larger the membrane area is the better) and concentration of removing agent in sample buffer
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