2 replies to this topic

[zelpop]

Does anyone know how to prepare HEPES-Tris Buffer.

I need to prepare 20mM HEPES-Tris Buffer, pH7.2. I found this buffer combination in one paper and thus I would like to test it.

The problem is: I don't know whether 20mM reffers to HEPES or Tris concentration? Or to both? And how to calculate in which ratio HEPES & Tris should be mixed?

Or I should simply make 20mM HEPES and adjust pH with Tris up to 7.2?

Thank you in advanced!

Zed

[protolder]

zelpop, on 28 January 2011 - 06:53 AM, said:

Does anyone know how to prepare HEPES-Tris Buffer.

I need to prepare 20mM HEPES-Tris Buffer, pH7.2. I found this buffer combination in one paper and thus I would like to test it.

The problem is: I don't know whether 20mM reffers to HEPES or Tris concentration? Or to both? And how to calculate in which ratio HEPES & Tris should be mixed?

Or I should simply make 20mM HEPES and adjust pH with Tris up to 7.2?

Thank you in advanced!

Zed

Hola, Tris, buffers betwen 7-9 and Hepes in 7-8. So for 7.2 you can use only one of them, but if you want to use the mixture with 20mM of each, you will have a more potent buffer capacity. Buena suerte

[zelpop]

protolder, on 30 January 2011 - 11:16 PM, said:

zelpop, on 28 January 2011 - 06:53 AM, said:

Does anyone know how to prepare HEPES-Tris Buffer.

I need to prepare 20mM HEPES-Tris Buffer, pH7.2. I found this buffer combination in one paper and thus I would like to test it.

The problem is: I don't know whether 20mM reffers to HEPES or Tris concentration? Or to both? And how to calculate in which ratio HEPES & Tris should be mixed?

Or I should simply make 20mM HEPES and adjust pH with Tris up to 7.2?

Thank you in advanced!

Zed

Hola, Tris, buffers betwen 7-9 and Hepes in 7-8. So for 7.2 you can use only one of them, but if you want to use the mixture with 20mM of each, you will have a more potent buffer capacity. Buena suerte

Thank you...Though I know these information and I was surprised that they used buffer mixture, maybe safer would be just to use HEPES...Thank you anyway for opinion