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Protein Standard not Appearing on Blots but Samples Do?
I've been having troubles with using Invitrogen's MagicMark XP Western Protein Standard. It seems that standard never appears when developing a chemiluminescent blot, even though all my samples do. No standard bands appear with a Ponceau S stain either. Only staining an extra standard with coomasie overnight shows any hint of bands. I've tried transblotting at 30 V overnight, 70 V for 4 hours, using different concentrations of standard, but nothing seems to work. My transfer buffer is 0.025 M Tris, 0.192 M Glycine, and 20 % MeOH, pH 8.3. Has anyone seen anything like this?
Re: Protein Standard not Appearing on Blots but Samples Do?
I think you should contact the manufacturer and suggest the batch is faulty. Must have been degraded, as you should see it with Ponceau S stain. Must be very little protein there.
|appearing , blots , protein , samples , standard|
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