I've tried loading various amounts of the BioRad Broad Range MW marker. Load too much, the whole lane is a smear. Load less, and only one band appears but it's way over-saturated. Google only gives me a discontinued product - is there a MW marker that shows up well with silver stain? Thank you!
I use Fermentas PageRuler™ Low Range Unstained Protein Ladder (SM1891), marker with 8 bands (3.4, 5, 10, 15, 20, 25, 30, 100 KDa) It works very well.
You might want to consider making your own using protein standards like Albumin, Ferritin etc with known molecular masses so that you can adjust their concentration individually to suit your particular application. You can do this by calibrating each protein standands with serial protein concentrations and seeing which concentration gives you the best result. Once you have each optimum standard concentration, just mix them in a master mix together and use as a normal protein ladder. It might take a little work to optimize at first, but once you have it you can freeze your master mix and use it indefinitely. I normally do this for my Blue-native gels and it has worked well. Good luck!
Unstained protein Marker of Bangalore Genie (merged to MERK Bio Science) also work very well.
Thank you all again -I tried the Prescision Plus Dual Color dilution, as we had some on hand, and it worked really well!
I may be late but if still facing the problem then you can try L/M/HMW calibration kit for SDS PAGE by Amersham Biosciences.
Hi All,
I had the exact same question as Jaclyn and have been using the following protocol which yields good results (see attached image).
Best of Luck!
-William
Diluted BioRad Precision Plus Dual Color standard 100X with 1x Laemmli sample buffer.
Loaded 10 µL of above into 20 µL wells of BioRad 12% Mini-PROTEAN® TGX™ Precast Protein Gels, 12-well, 20 µl #4561045.
Electrophoresed at 200 V (constant) for 38 min.
Fixed in 40% methanol, 10% acetic acid.
Stained with Silverquest staining kit from LifeTech (~3.5 minute development).
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