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FOCUS™ Extraction Buffers

Catalog Description Size Price(USD) Qty
786-219
FOCUS™ Extraction Buffer V For 50ml
$134.00
786-220
FOCUS™ Extraction Buffer I For 50ml
$90.00
786-221
FOCUS™ Extraction Buffer II For 50ml
$134.00
786-222
FOCUS™ Extraction Buffer III For 50ml
$134.00
786-223
FOCUS™ Extraction Buffer IV For 50ml
$134.00
786-233
FOCUS™ Extraction Buffer VI For 50ml
$134.00
786-234
FOCUS™ Extraction Buffers I-VI For 10ml Each Buffer
$234.00

FOCUS™ Extraction Buffers

Chaotropic extraction buffers that preserve the native charge of proteins making them suitable for sample extraction for 2D gels

 

One of the most important considerations before running 2D gel electrophoresis is the choice of protein solubilization buffers. A suitable buffer must solubilize proteins effectively, without disturbing the native charge of the proteins.  

 

Urea, a common chaotrope, is widely used for solubilization and denaturation of proteins. One of the disadvantages of using urea is carbamylation. In water, urea exists in equilibrium with ammonium cyanate, the level of which increases with increasing temperature and pH. Cyanate reacts with α-amino and ε-amino groups of proteins and induces a change in the isoelectric point of proteins. This leads to artifactual results. Therefore, carbamylation must be avoided. 

 

One way to minimize the risk of carbamylation is to prepare the urea-based reagents fresh before each use. G-Biosciences has developed a series of dry, urea-based, pre-mixed and ready-to-use solubilization buffers. Simply add an appropriate volume of the supplied rehydration buffer to the dry buffer. Then use to solubilize proteins, saving time and improving the quality of IEF/2D gel electrophoresis (Patent Pending).

 

FOCUS™ Extraction Buffers are also designed to be used as rehydration buffers for IPG strips. 

 

FOCUS™ Extraction Buffers are experimentally optimized concentrations of critical, buffering and stabilizing agents, including urea, thiourea, Nonidet® P-40, CHAPS and sulfobetaines. The FOCUS™ Extraction Buffers are designed to produce optimal protein extraction and improved spot resolution for 2D gel analysis. 

 

A range of FOCUS™ Extraction Buffers have been developed. Depending on the nature of the samples, one or more of the buffers suitable for your applications can be ordered. FOCUS™ Extraction Buffer-I is suitable for most applications; however, for stronger solubilization effects, we recommend FOCUS™ Extraction Buffer-II, -III, -IV, -V or -VI. A trial kit that contains all six FOCUS™ Extraction Buffers and each of their corresponding diluents is also available. The trial kit is designed for those who prefer to experiment for establishing a suitable extraction and solubilization buffer for their protein samples.

 

The following FOCUS™ Extraction Buffers, listed with their major components, are available: 


 
Major Components
FOCUS Extraction Buffer I Urea & Nonidet® P-40
FOCUS Extraction Buffer II Urea & CHAPS
FOCUS Extraction Buffer III Urea, thiourea, CHAPS & amidosulfobetaine (ASB)-16
FOCUS Extraction Buffer IV Urea, thiourea, CHAPS & sulfobetaine (SB) 3-10
FOCUS Extraction Buffer V Urea, thiourea & CHAPS
FOCUS Extraction Buffer VI Urea, thiourea, CHAPS & non detergent sulfobetaine 201

For analysis of a proteome, a single buffer may not be suitable and sequential solubilization using different FOCUS™ Extraction Buffers will help in identifying more and new proteins (see figure 1). See the application note in the technical section below for more information.

 

Features

  • Convenient & easy to use: simply hydrate and use
  • Prevents urea-induced protein carbamylation
  • Prevents waste of unused reagents
  • No artifactual protein bands due to dust & human skin contamination

 

Applications

  • Optimal protein extraction & solubilization for 2D gel electrophoresis & other applications
  • Rehydration buffer for IPG-strips
  • Improved spot resolution for 2D gel analysis

 

 

Protocol
786-219
786-220
786-221
786-222
786-223
786-233
786-234
Material Safety Data Sheet
786-219
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786-220
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
786-221
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
786-222
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
786-223
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
786-233
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
786-234
English_US
Danish
Dutch
English_UK
French
German
Spanish
Norwegian
Portuguese
Finnish
Swedish
Polish
Technical Literature
Improved Protein Extraction for 2D Electrophoresis The use of the FOCUS™ Extraction Buffers in sequential solubilization to identify novel proteins by 2D electrophoresis.
Mass Spectrometry Sample Prep Handbook A guide to the preparation of protein samples for Mass Spectrometry, including protein extraction, clean-up and peptide generation.
Plant Proteomics Handbook
Protease & Phosphatase Inhibitors & Proteases Handbook  A handbook & selection guide for inhibitors of protease & phosphatases & for proteases & assays
Protein Electrophoresis Handbook A guide to 1D and 2D protein electrophoresis products, including protein markers, electrophoresis buffers, 2D electrophoresis reagents, clean-up reagents and stains. The guide also offers protein sample preparation products.
Sample Preparation Handbook For Lysis Buffers, Fractionation, Dialysis, Protein Concentration and Enrichment
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