Hi
I found that there are two different recipes for transfer depending on the mode (Semi-dry or wet) of the transfer. Why is it so? If anybody has prior experience with the use of both the methods as well as recipes? Which one did you find more convenient and which one was better suited for your experiment? If you have image(s) as well please attach it as well.
Thanks
Hi, the choiche of wet or semy-dry depend on different elements. First of all what is your project and what proteins do you have to analyse? Semy-dray work better with proteins up to 70KDa, for high molecular proteins it is better the tank apparatus (wet)
Semy dry methods use less reagents (than tank apparatus) but it is more expensive (than tank apparatus) if you need to buy all the apparatus or some accessories.
So study your project to indentify the proteins that you need to study, find in your lab what type of equipment you have and what you need. Write a price list for reagents and equipment that you need and decide accordingly.
This site can help you but you can find a lot more on the web
https://www.licor.com/bio/guide/westerns/transfer_options
In my experience, the wet transfer is usually the way to go for wide range of proteins, time and economics of 1 system. I have succeeded from 6kDa to 200kDa with the Criterion system from Bio Rad for the past 20 years. Very easy system and very reliable for repeatable results. Tris/HCL gels, I prefer gradient gels as well, due to best separation. This system allows for a 40 min transfer so the whole run from start to finish can be completed in a day. I also like the 12+2 criterion gels they offer due to the ease of analysis in the end, the bands lay nice, easy to box in for analysis, and clean separation between wells. Cheers!
Well, I have to analyze proteins ranging from 21 KDa to 140Kda. I used semi-dry mode of transfer and found that it didn't perform the efficient transfer of 20KDa protein, however, there could be other reasons for it. One troubleshooting I did was increasing the methanol concentration to 20% and removing SDS from the transfer medium completely, however, it didn't have much impact on the transfer and that's when I found these two different recipes and it also just made me curious if anyone has had prior experience with both the recipes. Like Kellie said wet transfer is the more followed and convenient way to go for transfer of a wide range of proteins. Thanks for your help both of you.
- Badges
- Science topic
- Similar topics
- Computer Science
- Images