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NuPAGE bis-tris gel transfer buffer question


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#1 iridium

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Posted 08 February 2010 - 09:07 AM

If I want to transfer the NuPAGE bis-tris gel, can I use Tris-Glycine transfer buffer? Is it absolutely necessary to use the recommended NuPAGE bis-tris transfer buffer?
Another question is: If I use the 4xLDS sample buffer to prepare my sample, can I run it on a regular Tris-glycine gel? Thank you!!

#2 Lyanesse

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Posted 11 February 2010 - 01:00 PM

In our lab (where we run probably 50 westerns a month), we regularly transfer NuPAGE bis-tris gels with Tris-Glycine transfer buffer, and have excellent results. I have used both the NuPage transfer buffer and Tris Glycine, but prefer the Tris Glycine actually. We use TG transfer buffer from Fisher with 20% methanol, transfer at 35V for 1.5hr in a large BioRad transfer apparatus. We like the larger tank so we can transfer midi gels or multiple mini gels. We also re-use the TG buffer for up to 3-4 months with no detectable change in transfer efficiency, so this saves $$ compared to the Invitrogen protocol making fresh transfer buffer every time.

I haven't tried running TG gels with the LDS buffer, so can't help you there.

Good luck!

If I want to transfer the NuPAGE bis-tris gel, can I use Tris-Glycine transfer buffer? Is it absolutely necessary to use the recommended NuPAGE bis-tris transfer buffer?
Another question is: If I use the 4xLDS sample buffer to prepare my sample, can I run it on a regular Tris-glycine gel? Thank you!!






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