Hi, I run two 2D gels electrophoresis using a defined pollen extract and protocol with an interval of several months. In both cases, I load equal amount of protein. Unfortunately, results of 2D are different (images of two 2D gells to be attached, image 1 is the first gel). What cause this problem? Could this be due to strips? I used BioRad strips 3-10 NL, 18cm).
Thanks!
protein load: 400 µg
pollen proteins defatted using acetone, extracted with PBS (buffer phosphate saline) and dialysis against distilled water.
I dont think it is due to strips. Horiziontal streaking could be due to many problems like, improper sample preparation leading to high salts, nucleic acids, polysacharide contaminations. Please find the attached document for your reference.
The striking in the gel is looking mostly due to salts. Please refer the GE manual for trouble shooting guide. I am attaching the same for your reference.
the problem is in the first dimensions, maybe the samples is not the same, and your buffer, you can use a kit too clean your sample from salts
thanks to all, the samples are the same, but for samples 2, rehydration buffer is lack of carrier ampholytes. How can I improve the quality of the pollen extract? What method do you recommend for extraction?
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