I purchased this Merck millipore's DURAPORE PVDF 0.45 micron WH PL 13 mm hydrophobic filter instead of 0.45 um syringe filter. Now I am wondering if it is suitable to use it for purifying my intracellular protein? If yes, how do I make the assembly of it and how to reduce the chances of contamination?
I suppose that you have a filtering device into which you can insert the filter with the proper gasket to ensure that liquid is not going to leak out from the side when you force liquid through it with a syringe. Then, just mount the divice with the filter inside, but without tightening the gasket, wrap in aluminium foil and autoclave. After autoclaving, remover the aluminium foil, tighten the gasket and use the divice to filter sterilize your protein (to be done with the purified protein, of course, to avoid contamination in further purification steps)
Hi Sailee Asolkar, Dr. Pierre Béguin gave a nice option to sterilize. You can also use BSL to reduce more contamination. The filter you are going to use should be fit in the filter holder it shouldn't be smaller to prevent leakage. Then you can have a purified sample
Hi Dr. Pierre Béguin thank you for the answer. The "filtering device" is what I am looking for. It would be great if you recommend the best ones, especially for a 13 mm filter.
Thank you Srichandrasekar Indhuprakash for the inputs. I do have access to BSL and shall proceed with that. Also about the filter holder, kindly suggest the make of the ones you use for routine filtering processes.
Sailee Asolkar Filter holders are available in Tarson's (we are using this one) and Merck make, you can choose your desired filter holder based on the size and need.
If you pass protein solution over a PVDF membrane, all protein will bind to it non-selectively (well, at least until the membrane is saturated). I am therefore not sure how this would contribute to purification.
Some 30 years ago I used a custom-made filter holder to do non-equilibrium kinetics with an enzyme immobilised on PVDF (because syringe-filters with this membrane were not commercially available at the time), private-mail me for the drawings if you are interested.
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