Hello
I am planning to perform SDS Gel electrophoresis and my protein is about 6kb. I was checking on BioRad and saw many kinds of precast gels.
1) How should i decide whether to use Tris Hcl or Tris-Acetate or TGX or Tris Tricine gels? What would change according to these?
2) I think I need to use mini protean or criterion? Which of them (mini protean and criterion) would be better for my protein in your experiences? I couldnt see a the difference other than the gel size.
3) Anyone ever used precast gels? Yay or nay? Who do you think is the best precast gel maker?
Thanks a lot!
Pricing, gel shelf life, pre-made buffers, versatility, and apparatus are considerations as well but Novex gels seem to be more widely used in industry (bulk pricing) from what I've seen. There are usually charts from the various vendors to show the effect of different gels and their buffers on the separation of the desired protein from the contaminating proteins. So the type of gel may depend upon the contaminants that can migrate closely to your protein. Precast is convenient and consistent but at a price..
I like using the precast 4-20% TruPAGE gels from Sigma because they are difficult to tear. With the MOPS running buffer, your 6 kDa band should be on the gel.
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Bulletin/2/pcg3001bul.pdf
I use Bio-Rad criterion sized gels and prefer the TGX form as they have very long shelf lifes and work with the standard Tris-glycine (Towbins) buffer. It seems a lot of folk use the mini gels, but the larger midi format allows one to spread similar weighted proteins further apart if you happen to need that. So far I've never had a bad pre-cast gel from Bio-Rad. The Bio-Rad "stain free" gels can be useful if you are wanting to image your gels. They are of little use for imaging a blot, however, unless you have an imager that has epi illumination in the UV range. They do work and makes it easy to quickly assess the gel before proceeding. A lot of folk use Novex gels and equipment too.
Hmmm. Yes for me long shelf life is important. I should be able to view small kd protein and also prefer a hard to tear gel.
Thanks for valuable inputs. I will check sigma aswell. I checked Novex but it looks like their cassettes are harder to work with when compared to BioRad TGX. Bio rads wells seem easier to load aswell.
By the way is it possible to use Sigmas BioRads or Thermos cassettes with the other vendors equipment? Would they fit as long as the machine and the cassette are both mini or midi sized ?
Sigma's 10 x 10 cm TruPAGE gels fit the Novex minigel box perfectly.
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