dissolving of SDS-PAGE bands

ottcr at esvx17.es.dupont.com ottcr at esvx17.es.dupont.com
Mon Jun 21 17:16:56 EST 1993


In article <1993Jun18.011037.9650 at anat.UMSMED.EDU>, jbh at anat.UMSMED.EDU (James B. Hutchins) writes:
>
>In article <1vpduiINN1ukm at rs1.rrz.Uni-Koeln.DE>
>thomas at biomed.biolan.uni-koeln.de writes:
>>Hi,
>>has anyone a reliable protocol for dissolving of SDS-PAGE protein bands for 
>>  >>>ETC. ETC. (STUFF DELETED)
>>
>>Thomas
>>
>
>Unfortunately, I'm at home and do not have the reference, but having done
>gel slices too many times :-( I remember the formula by heart:
>
>99 ml 30% H2O2          -
>1 ml conc. NH4OH
>Heating to 37 deg C (or even 60 C) is sometimes required.  We drop the slice
>directly in the scintillation vial, place them loosely capped in a large
>37 deg C incubator overnight, and add scintillant, cap, mix, chill and count.
>

FIND REFERENCE : Mahin and Lofberg; Anal. Biochemistry; 16, page 500 (1966)
   This reference is similiar to the above. It also includes perchloric acid.
   Both methods, given the strong oxidizing agents, can loose sample
   (exploding vials/caps). Both methods will produce chemiluminescence
   (extreme). 
   A common technique for the past 10 years has been elution using commercially
   available solubilization reagents. With the elution technique, approx. 82%
   or better recovery can be obtained, which is only slightly less than the 
   Mahin Lofberg technique. 

For more info send me E-Mail
Regards, Charly & Rich



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