i am working with sap proteins which were not visible on SDS-PAGE, i was just able to see the band by using Tris Tricine Gel. Now i am confused if i use SDS-PAGE in 2D will i be able to see the proteins? kindly guide,
thankind in advance
If your proteins are not visible in 1-D SDS-PAGE, why should they become visible in 2-D SDS-PAGE ? If you need 2-D gel analysis, you should adapt the technique and try Tris-Tricine in the second dimension (I suppose that the first dimension is IEF, as usual).
Sheher Yar
The question is how did you visualize your SDS-PAGE gel ? I do not see a problem using Tris-Glicine in 2D electrophoresis. I have done it in the past but as Pierre Béguin already mentioned I have no idea how it should increase visuablitiy of your band...
Have you considered using more sensitive staining of your gel or loading more material ?
Pierre Béguin thank you so much sir for your time and precious suggestion, i am a little bit confuse that if i will use the spacer gel also in 2D (in the case of Tris Tricine) or i will just use the seperating gel only? i am sorry if the question is very ordinary.
I would try the same Tris-Tricine buffer system as you did for the 1-D gel, including the spacer and the separating gel. However, the problem may be equilibrating the gel of the first dimension with the sample buffer of the second dimension, while preventing small polypeptides from leaching out of the gel
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