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Sucrose Gradient Analysis of Proteins

Hui Zhu Hui Zhu

Published: Feb 5, 2012 DOI: 10.21769/BioProtoc.194 Views: 30336

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Abstract

The sedimentation rate of a protein in a linear sucrose gradient can be used to determine its S value when compared to proteins of known S values which are run in a parallel gradient. This can be used to estimate an approximate molecular weight of the protein and test the potential interaction of two proteins if they peak in the same gradient range.

Keywords: Sucrose gradient search Molecular weight of the protein search S value search

Materials and Reagents

  1. Sucrose (Sigma-Aldrich, catalog number: S0389 )
  2. Gelatin (Sigma-Aldrich, catalog number: G9136 )
  3. Aldolase (Bio-Rad Laboratories)
  4. Thyroglobulin (Bio-Rad Laboratories)
  5. Sucrose solutions (see Recipes)

Equipment

  1. Centrifuges (Beckman Falcon, TLS-55)
  2. Gradient mixer (Sigma-Aldrich)
  3. Stirring bar

Procedure

Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.

Category

Biochemistry > Protein > Quantification

Cell Biology > Cell-based analysis > Transport

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10% is 10 g/100 ml40% is 40 g/100 ml
edit 1 Answer 4 Views Feb 19, 2013