I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band.
I was thinking of using a standards ladder or make some standards are different concentrations and compare my band to it.
2 things:
1) does anyone have a protocol they could please share with what software they use etc
2) Is it possible that this can be done thought a normal printer scanner instead of a fancy GelDoc?
Thanks.
Hi Akash,
If you prepared standards of known protein concentrations, you can use our Phoretix 1D software to extrapolate the protein concentration within your bacterial lysate under the "quantity calibration" step.
You can get a free trial of Phoretix 1D from our website here: https://totallab.com/products/phoretix-1d/
Yes you can use a normal scanner to scan your image rather than a gel doc, however make sure that you use the highest quality image capture settings for best results - we recommend an image of at least 16-bits and to use the .tiff file type for export.
Best wishes,
Steven
Densitometric analysis can be done by Imagelab software, which is a free software. Please go through the protocol of that software. I recommend to do Gel doc pictures as they are of high resolution (600 dpi) ( in tiff format).
Hi Akash,
It is more convenient to use Biorad's ImageLab program (as noted above, it is free). There are tutorial videos on Biorad's YouTube channel on how to do this. The latest version of the program allows you to process .tif files (gray scale 8 bit) from other sources (but there may be problems with image settings, not all are simply imported). Or as an option - the free program ImageJ, there are a lot of video tutorials on it too
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