Slimy protein lysates
Posted 26 January 2009 - 06:20 AM
after using this protocol for the extraction of nuclear and cytosolic proteins I sometimes end up with very slimy lysates after heating them up (95°C for 5') before putting them on a gel. This is really putting me off..since I've tried amost everything to avoid this. i.e. avoiding to high protein concentrations in the lysates, spinning them down @ 13.000 rpm for 15 minutes before protein quantification and before loading the gel to avoid transfer of sediments...I'm running out of ideas...
Advice would be highly appreciated!
Posted 26 January 2009 - 01:32 PM
Posted 27 January 2009 - 02:15 AM
Posted 19 February 2009 - 01:45 PM
Thank you! You're right! It's mostly the nuclear lysates that are causing the problem... but what can I do?? Use DNAse? What can I do to make samples easier to load after boiling? Maybe spin them down longer and will the chromatin be in the pellet after that?
you can try using a syrynge and needle, pull sample up and down a couple of times and then try again.... hope this helps