I'm trying to do total protein quantification with Stain-free precast gels and after a lot of reading and many phone calls with Bio-rad, I am confident it's possible to read the gels and membranes with just UV box or an imager with UV capabilities. Has anyone done this before? How have you been able to get good images and measurements? So far, using a G-Box imgaer has given me pretty bad pictures, but I can see the bands so much clearer on just a regular old UV transilluminator box in my lab. Any suggestions for getting better images for normalizing total protein?
I am not sure what are software options for G-box imager, but if we on old transiluminator with mounted photo chamber can shorten exposition time, there must be an option like that for newer devices. We also had problems when camera that refused to focus so we changed gel height by putting glass plates under gel ( in the end it was temporary software glitch)
Hi,
maybe this publication could help:
https://doi.org/10.1016/j.ab.2003.10.047
Btw, you could also "stain" standard SDS-PAGE gels by soaking in TCE after running the gel, but then there is not much of an advantage left compared to other staining methods. The method also works with Trichlor-acetate (only after the run, because TCA can't be added to the gel directly before the run due to its charge).
I guess the imaging system has been optimized in terms of wavelength, exposure time and image processing so if you can afford it, you can save some time. Or spend some time at a regular imager and optimize yourself.
Hope that helps.
Once i transfer protein to membeane ns and wet with methanol to get a transilumination effect. I could see the total protein around the wet methanol. Then could do western Immunostaining.
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