[RASMB] proteins in water

[Lake Paul]

Carlos,

If your protein is charged then you will definitely need some counter ions
in there. That is the most obvious cause for lower than expected
sedimentation coefficients. Here is a good paper to start with design,
buffer considerations etc etc.

Hope this helps, 

Lake

Analytical Ultracentrifugation in the Study of Protein Selfassociation

and Heterogeneous Protein-Protein Interactions 

Andrea Balbo and Peter Schuck

 

Lake N. Paul, PhD

Biophysics Research Specialist

Discovery Park - Bindley Biosciences Center Purdue University

1203 West State Street

West Lafayette, Indiana, 47905

765.494.4960

BioAnalytical Lab Website:

http://www.purdue.edu/discoverypark/bioanalytical/

 

From: rasmb-bounces at rasmb.bbri.org [mailto:rasmb-bounces at rasmb.bbri.org] On
Behalf Of Carlos Alfonso Botello
Sent: Friday, July 20, 2012 11:17 AM
To: rasmb
Subject: [RASMB] proteins in water

 

Dear colleagues,

We are running a 10 kDa protein at concentrations 0.2-2 mg/ml in water (no
buffer is present).The sedimentation coefficients that we are obtaining are
lower than expected for the monomer (1.4 - 1.5S). Is there any correction of
the S that we could apply to obtain the correct S20,W.

Thank you in advance for your help,

Carlos





Dr. Carlos Alfonso
Departamento Biologia Celular y Molecular
Laboratorio de Interacciones Macromoleculares: Bioquímica Citomimética
Centro de Investigaciones Biológicas, CSIC
C/ Ramiro de Maeztu, 9. 
28040-MADRID
Tlfno.- 91 8373112 ext.4406-4408-4297 
Fax: 915360432.
E-mail: carlosa at cib.csic.es
NEW lab web page: 
http://www.cib.csic.es/es/grupo.php?idgrupo=49
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