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Protein Standard not Appearing on Blots but Samples Do?

Protein Standard not Appearing on Blots but Samples Do? - Western Blot Forum

Protein Standard not Appearing on Blots but Samples Do? - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 01-11-2011, 09:08 PM
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Default Protein Standard not Appearing on Blots but Samples Do?



Hi all,
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?

Thanks
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Old 02-18-2011, 12:14 AM
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Default 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.
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