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HELP NEEDED: How to prepare HEPES-Tris Buffer?


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#1 zelpop

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Posted 28 January 2011 - 06:53 AM

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

#2 protolder

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Posted 30 January 2011 - 11:16 PM

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

#3 zelpop

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Posted 31 January 2011 - 01:58 AM


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




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