Selection of membrane is very important for expression of proteins with good quality in WB. So I want to know the best membrane and its supplier so that good results can be obtained.
PVDF membrane from Millipore is very good. My lab colleagues have been using it for a long time and it is working very well.
https://www.emdmillipore.com/US/en/product/Immobilon-P-Membrane%2C-PVDF%2C-0.45%C2%A0µm%2C-26.5%C2%A0cm-x%C2%A03.75%C2%A0m-roll,MM_NF-IPVH00010
Hello
You can Nitrocellulose membrane from Life technology
regards
PVDF membrane from Millipore is very good. My lab colleagues have been using it for a long time and it is working very well.
https://www.emdmillipore.com/US/en/product/Immobilon-P-Membrane%2C-PVDF%2C-0.45%C2%A0µm%2C-26.5%C2%A0cm-x%C2%A03.75%C2%A0m-roll,MM_NF-IPVH00010
I would also upvote for Millipore PVDF and we had terrible experience with GE healthcare membrane(would rate poor).
PVDF membranes from Biorad for big proteins. however if you are transfering a small protein (ie. 10 Kd>) then use nitrocellulose from biorad
Big proteins mean... My proteins of interest are within 12-100 Kd range.
There are actually quite a few variables that can affect the results of a western and influence the membrane of choice. As Fabrice noted, both can be utilized with excellent results. Your transfer efficiency for larger and smaller proteins will depend on the pore size, regardless of the membrane you use. Pall makes several different pore size for both NC and PVDF, and I've had success with both. Where PVDF does have a slight edge is when you have a large amount of background, as PVDF does a better job of eliminating background, in my opinion anyway. My rule of thumb, for new antibodies, I try both and see which works better, as I don't believe there's a hard and fast answer to your question, but I do agree with the brands suggested.
Thank you all for your valuable suggestions and sharing of your experience related to membranes for WB.
Hi Vinay, as others have said, PVDF is usually a good first choice (be sure to soak it in MeOH briefly first before use, as recommended by the manufacturers).
It does depend on your proteins, too. Hydrophobic proteins will be captured much more efficiently by PVDF than nitrocellulose (which may be the reason they apparently "give less background" with PVDF); the reverse will probably be true if your proteins are strongly hydrophilic.
The pore size is also important to bear in mind as you may need to go for a smaller size than 0.45 µm to get good results with proteins as low as 12 kDa. In other words, if you're ever having trouble seeing the bands with your smaller proteins, try a smaller pore size membrane (0.2 µm). Hope that helps!
Millipore have a good portfolio of PVDF membranes, which fit well with their work flow: if you plan to make tonnes of blot their workflow is worth the investment (precast gel + transfert + turbo detection)!
If you do not plan to buy their complete system, GE put a lot of effort to propose a good and complementary portfolio of great membranes (including various nitrocellulose/PVDF optimized for many different applications). Contact them they will help you to choose what you need.
If to complicated, the biorad 0.2µm nitrocellulose not supported is a wide standard in many many many lab. Work correct for almost everything.
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