3 replies to this topic

[jasmina]

Hi,
  I want to run a gel SDS PAGe to do WB on my protein of 55kda.
I m wondering what could be better if do 10 or 12% gel acrylamide. since I would perform western blots of actin and my protein in the same membrane, but successively. without cutting the filter.
I already tried 10% gel, I saw my protein ,but after I couldnt separate well actin and my protein molecular weights, particulary for further quantification with image J.
thanks for advice

[protolder]

Hola, I think can be better 8% than 12%, actin could run near the front but the resolution could be better. Buena suerte

[jasmina]

protolder, on 14 March 2011 - 10:51 PM, said:

Hola, I think can be better 8% than 12%, actin could run near the front but the resolution could be better. Buena suerte

to separate better vgat from actin bands, 12% is good? since i found it in one paper!

[protolder]

luciana, on 24 March 2011 - 07:47 AM, said:

protolder, on 14 March 2011 - 10:51 PM, said:

Hola, I think can be better 8% than 12%, actin could run near the front but the resolution could be better. Buena suerte

to separate better vgat from actin bands, 12% is good? since i found it in one paper!

Hola, as you know concentrated gels are suitable for separation of small proteins and diluted for big ones . probably you have the reason (Im sorry) but all the coloured mw markers have protein in your interval. Look for some markers in your lab or institution and run two gels to test. I repeat my excuses if I confuse you. Buena suerte