Roti-Load versus original Laemmli sample buffer and temperature

11 November 2008 1 3K Report

Hi,

could I start a thread on an improved Laemmli buffer. Roti-Load is based on several papers showing that with phosphate buffer and lower temperature (65°C rather than 95°C) one gets less protein degradation. There is no need to boil Laemmli anyways; some transmembrane proteins can actually form aggregates.

What's your experience.

Harald

Harald H.H.W. Schmidt

Hi Nico,

thanks for looking into this. The 3 relevant papers that I found are:

(1) Cannon-Carlson, S. and Tang, J. (1997), Anal. Biochem. 246, 146-148.

(2) Kowit, J.D. and Maloney, J. (1982), Anal. Biochem. 123, 86-93.

(3) Rittenhouse, J. and Marcus, F. (1984), Anal. Biochem. 138, 442-448.

Carl Roth AG (Germany) sells Roti-Load R.

Proteins denature at 65°C, so I never understood why 95°C should be necessary.

Looking forward to hearing from you.

Cheers,

Harald

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