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2D electrophoresis

The two dimensions that proteins are separated into using 2D electrophoresis are isoelectric point and protein mass (molecular weight).

To separate the proteins by isoelectric point is called isoelectric focusing (IEF). Thereby, a gradient of pH is applied to a gel and an electric potential is applied across the gel, making one end more positive than the other. At all pH values other than their isoelectric point, proteins will be charged. If they are positively charged, they will be pulled towards the more negative end of the gel and if they are negatively charged they will be pulled to the more positive end of the gel. The proteins applied in the first dimension will move along the gel and will accumulate at their isoelectric point; that is, the point at which the overall charge on the protein is 0 (a neutral charge).

After completion of the first dimension the complexes are destroyed by applying the denaturing SDS-PAGE in the second dimension, where the proteins of which the complexes are composed of are separated by their mass. The result of this is a gel with proteins spread out on its surface. These proteins can then be detected by a variety of means, but the most commonly used stains are silver and Coomassie Brilliant Blue staining.

The final goal most often is to determine each protein spot from the 2D gel most often via MALDI analysis, for this spot picking of the proteins out of the 2D gel (manual or automated), removing the spot from the gel, digest it and treat it to be ready to spot on a MALDI target is the workflow to follow.

Product namecontents
Hoefer IEF100
Mini 2-D Electrophoresis SystemIEF-100
Mighty Small II
Spacers
Standard 2-D Electrophoresis SystemIEF-100
Chroma
Power supply
Large 2-D Electrophoresis SystemIEF-100
SE900 Large Unit
Hinged glass cassettes
Multiple gel caster
Large 2-D Electrophoresis system w/ power supplyIEF-100
SE900 Large Unit
Large hinged glass cassettes
Multiple gel caster
Power supply
 

Proteomics

ProPrep LC
IMA transfer plate
 

Protein dyes

FluoroProfile protein quantification kit
LavaBlue protein stain kit
LavaCell Live imaging stain
LavaPep - peptide quantification kit
LavaDigest protease monitoring kit
LavaPurple total protein blot stain
Visio
 QPRO – BCA kit Standard
 QPRO – BCA kit Micro