Help with large SDS-PAGE gels

Brian J. Baumgartner brianb at bcm.tmc.edu
Thu Jun 30 14:22:21 EST 1994


In article <2us8dj$sd7 at nic.umass.edu>, AAT at UCSVAX.UCS.UMASS.EDU (Gus Trautweiler) writes:
> Receiving very poor resolution for mw proteins less than 25000 for gels 22cm x
> 18cm.  Smaller gels (12cm x 8cm) work.  Stacking gel
> conc.= 5% and tris pH6.8, running gel conc. have been run at 12%, 15%, 20% and
> tris pH8.8. Voltage was 100 stepped to 200 with milliamps at 30 maximum.  Total
> time of run approx. 5hr.  Thank you for your help, Timothy J. Byrne

Timothy,

You didn't say what your %C is, try 5-6% to improve resolution of lower MW
proteins.  Alternatively, try the Tricine-SDS-PAGE system.  They take
forever to run (~24 hr) but give beautiful resolution in your size range. 
Sigma and Promega sell MW markers for these gels.  The reference is: H.
Schagger and G. von Jagow, Anal. Biochem. 166, pp. 368-379 (1987).
Good Luck!

Brian Baumgartner
brianb at bcm.tmc.edu




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