The composition of Elution buffer - I want to know what is the composition of your buf (Jul/14/2005 )

Hi, all

Have someone use HEPES buffer in His-tagged protein purification

I want to know what is the composition of your buffer and how much Molar of each recipes ?

for example 50mM HEPES, 250mM NaCl, 500mM Imidazole.

Thank you

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I have used HEPES buffer for purification.
Solubilisation buffer: 20mM HEPES
300mM NaCl
20% glycerol

Wash buffer: 20mM HEPES
20mM imidazole
600mM NaCl
10% glycerol

Elution buffer: 20mM HEPES
200mM Imidazole (but have had problems eluting at this conc therefore suggest increasing this conc).
10% Glycerol
The pH of your buffers will depend upon the PI of your protein.
Hope this helps.

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