Im using 2x laemli buffer at 1:1 to load 35ug per well. But i have too low a concetration to get a desirable amount for each well. I already have the lysates and im on a time restriction so iwas wondering if i can increase the concetration of laemli buffer or something.
You can reduce the amount of laemmli buffer a little bit... like maybe 10-20% below the proper amount and it usually still works fine. I would not do this if directly comparing to other blots that were loaded with the usual amount. Also you can buy 4X laemmli buffer.
Hi there,
One possibility is to precipitate protein (TCA or acetone) and resuspend them directly in the LB.
You might consider a 5x loading buffer such as Pierce™ Lane Marker Reducing Sample Buffer. Also if you can borrow a Bolt chamber and 10-well Bolt gels you can load up to 60 ul volume per well.
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