I want to purchase the prestained protein marker for the WB for my proteins of interest (size of my proteins vary from 12-100 kD). So, can you suggest me the company which provide the good quality marker. I have very bad experience with the SRL, not satisfactory with the Thermo Scientific. Although i have found the good result with fermentas. What about the prestained markers from Biorad or some other
We have been using prestained markers from Biorad. You can go for the same :)
As Marta suggested Dual Color of biorad works very fine but finally you have always to take in mind that acrylamide percentage can influence even marker run. if you need to size your protein I suggest to use other two antibody ( an upper an a lower molecular weight) to detect the weight of your protein without any doubt.
regards
Francesco
I have used them all and feel no big difference. But, do remember when the dye is red/pink/yellow the color tend to fade away very quickly during wash. I am kind of going back to all blue since the color stays longer.
Hi, as a few other people have said, BioRad's markers have always worked fine for me (single or dual-colour, depending on how much you want to spend). The only brand I've tried which was unsatisfactory was NEB (very diffuse bands).
Best Regards,
Mark
Thanks for your valuable suggestions
Can you tell me the that how much volume (minimum vol) of Precision plus protein Kaleidoscope standards from BIO-RAD should be loaded in midi gel.
Dr. Kant,
I think you should consider the gel system that you use prior to considering the marker. If you use Invitrogen (or other pre-poured gel and buffer systems) pre-poured gels and buffer systems, their markers are very particular on which gel system. If you pour your own gels, I would start with biorad. Francesco has good suggestion to use antibody to molecular weight marker to be absolutely sure where the protein migrates. If WB is too much, stain proteins with Coomassie blue or Sypro ruby (fluorescent) and sequence MWM with MS. Also likely going overboard, but the markers migrate differently depending upon gel and buffer system.
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