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2D Gel Electrophoresis

Two-dimensional gel electrophoresis is a technique used to analyze complex protein mixtures. It involves two steps - first, isoelectric focusing separates proteins by their isoelectric points. Second, SDS-PAGE separates them by molecular weight. This results in thousands of protein spots on the gel corresponding to different protein species, and provides information on each protein's properties.

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132 views2 pages

2D Gel Electrophoresis

Two-dimensional gel electrophoresis is a technique used to analyze complex protein mixtures. It involves two steps - first, isoelectric focusing separates proteins by their isoelectric points. Second, SDS-PAGE separates them by molecular weight. This results in thousands of protein spots on the gel corresponding to different protein species, and provides information on each protein's properties.

Uploaded by

Nandhini D P
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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TWO-DIMENSIONAL GEL ELECTROPHORESIS

INTRODUCTION

Two-Dimensional Gel Electrophoresis (2-DGE or 2-D electrophoresis) is a powerful and widely


used method for the analysis of complex protein mixtures extracted from cells, tissues, or other
biological samples. Two-dimensional electrophoresis was first introduced by P. H. O'Farrell and
J. Klose in 1975. This technique sorts proteins according to two independent properties in two
discrete steps: the first-dimension step, isoelectric focusing (IEF), separates proteins acording to
their isoelectric points (pI); the second-dimension step, SDS-polyacrylamide gel electrophoresis
(SDS-PAGE), separates proteins according to their molecular weights (Mr, relative molecular
weight). Each spot on the resulting two-dimensional array corresponds to a single protein species
in the sample. Thousands of different proteins can thus be separated, and information such as the
protein pI, the apparent molecular weight, and the amount of each protein is obtained.
MATERIALS

1. Lysis Buffer

2. Rehydration Buffer

3. Equilibration Buffer

PROCEDURE

1. SAMPLE PREPARATION

2. ISOELECTRIC FOCUSING/THE FIRST DIMENSION

3. EQUILIBRATION

4. RUNNING THE SECOND DIMENSION

5. GEL SILVER STAINING

6. IMAGE AND DATA ANALYSIS

RESULT:

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