HELP NEEDED: How to prepare HEPES-Tris Buffer? - (Jan/28/2011 )
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
-zelpop-
zelpop on Fri Jan 28 14:53:11 2011 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
-protolder-
protolder on Mon Jan 31 07:16:23 2011 said:
zelpop on Fri Jan 28 14:53:11 2011 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
-zelpop-