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Hi,
Has anyone encountered protein degradation on 2D gels before?
I have sonicated my samples for 1 min (using a relatively mild blast-setting, chilled on ice, sample dissolved in urea/thiourea-based lysis buffer).
I applied the identical sonication settings for all my samples. Yet, I observed specifically that in all my 3 biological replicate control gels, protein spots were missing at the high MW, basic end (i.e. ~ 40-66KDa, pH 4-6). Nonetheless, the acidic, high MW proteins were not affected in these control gels, whilst at the same time there were no significant increase in small MW proteins (a sign of fragmented or degraded proteins).
This situation was not observed in gels from other treatments.
How would you interpret these results? Please give me some suggestions, thanks!
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White bands on Cy3 gels
14 December 2012 - 12:38 AM
Hi!
I imaged Cy3 gels using the Typhoon scanner coupled with a proper set of Laser, Excitation and Emission filters.
Yet, in addition to the conventional 'dark' Cy3 bands, I also observed that certain protein bands appeared as 'white' on the same gel.
Have you encountered this before? Or what may be the possible cause of this?
P.S. I have imaged the same gel using another fluorescent scanner (Pharos FX; Bio-rad). The 'white' bands still persisted, suggesting that it was likely not the scanner problem.
Thanks a lot in advance! Please kindly give me some advise.
I imaged Cy3 gels using the Typhoon scanner coupled with a proper set of Laser, Excitation and Emission filters.
Yet, in addition to the conventional 'dark' Cy3 bands, I also observed that certain protein bands appeared as 'white' on the same gel.
Have you encountered this before? Or what may be the possible cause of this?
P.S. I have imaged the same gel using another fluorescent scanner (Pharos FX; Bio-rad). The 'white' bands still persisted, suggesting that it was likely not the scanner problem.
Thanks a lot in advance! Please kindly give me some advise.
