ReadyPrep Sequential Extraction Kit

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Overview

Differential Solubility
Differential Solubility

The ReadyPrep sequential extraction kit is used to extract proteins of differing solubility from cell lysates. Based on the work of Molloy et al. (1998) and Herbert et al. (1998), the kit uses a series of three extraction buffers, each differing in their detergent and chaotrope concentrations (Rabilloud et al. 1997). The insoluble residues from each extraction step are further extracted with reagents of increasingly stronger solubilizing power.

The ReadyPrep sequential extraction kit:

  • Yields fractions that are suitable for direct analysis by a number of methods, including 2-D electrophoresis
  • Can be carried out in a single microcentrifuge tube, minimizing protein loss
  • Results in manageable 2-D protein patterns for simplified protein identification and gel matching
  • Generates fractions with high protein loads to enable visualization of low-abundance proteins

 

References
Molloy, MP, et al. (1998). Extraction of membrane proteins by differential solubilization for separation using two-dimensional gel electrophoresis. Electrophoresis 19, 837–844.
Herbert, BR, et al. (1998). Improved protein solubility in two-dimensional electrophoresis using tributyl phosphine as reducing agent. Electrophoresis 19, 845–851.
Rabilloud, T, et al. (1997). Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 18, 307–316.

ReadyPrep Sequential Extraction Kit

163-2100
5–15 preps, protein preparation and fractionation kit, for differential solubilization with SB 3-10, includes 3 premixed reagents, TBP, 3 protocols

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ReadyPrep Sequential Extraction Kit Reagent 1

163-2102
50 ml, 40 mM Tris base, for protein preparation and fractionation buffers; component of kit #163-2100

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ReadyPrep Sequential Extraction Kit Reagent 2

163-2103
10 ml, protein preparation and fractionation reagent, includes 8 M urea, 4% CHAPS, 40 mM Tris, 0.2% Bio-Lyte 3/10 ampholyte; component of kit #163-2100

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ReadyPrep Sequential Extraction Kit Reagent 3

163-2104
10 ml, protein preparation and fractionation reagent, includes 5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10, 40 mM Tris, 0.2% Bio-Lyte 3/10 ampholyte; component of kit #163-2100

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Number Description Options
4006167 Instruction Manual, ReadyPrep Sequential Extraction Kit, Rev B Click to download
3096 Sample Preparation: Tools for Protein Sample Extraction, Cleanup, Fractionation, and Depletion Brochure, Rev B Click to download [ Add to Cart (Free) ]