Laemmli Lysis-buffer
Protein isolation Bacteria - Bordetella pertussis
Publication protocol
Bacteria were grown in liquid Stainer-Scholte medium for 18 h at 37°C, and cells from 1 ml of bacterial suspension were collected by centrifugation (10 min, 15,000 × g). The pellet was resuspended in 100 μl of Laemmli buffer (50 mM Tris [pH 6.8], 2% SDS, 0.1% bromophenol blue, 10% glycerol, 1% β-mercaptoethanol) and dissolved for 5 min at 100°C. Ten microliters of lysates was separated by 7.5% SDS-polyacrylamide gel electrophoresis (PAGE) and transferred to a nitrocellulose membrane.Full paper Login or join for free to view the full paper.
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Discussion
Discussion
4 years ago
Author:
Hollie Fowler
How can I deal with my pellet being too viscous?
I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?
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