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Related Concept Videos

Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as  cells...
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Gradient Fields01:27

Gradient Fields

A gradient field is a vector field derived from a scalar field. A scalar field assigns a single numerical value to every point in space, such as temperature, pressure, or electric potential. The gradient field describes how that value changes from point to point. It gives both the direction of the fastest increase and the rate of change in that direction.For a scalar field f(x, y), the gradient is written as\begin{equation*}\nabla f=\left\langle \jfrac{\partial f}{\partial x},\jfrac{\partial...

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Related Experiment Video

Updated: Jun 14, 2026

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis
11:38

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis

Published on: February 29, 2016

Pouring linear gradient gels with a gradient former.

Richard J Simpson

    Cold Spring Harbor Protocols
    |April 3, 2010
    PubMed
    Summary
    This summary is machine-generated.

    Linear gradient gels offer superior protein separation compared to fixed-concentration gels by accommodating a wider range of molecular weights and resolving similar protein sizes more effectively. This protocol details their preparation for enhanced biochemical analysis.

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    Published on: February 12, 2013

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Proteomics

    Background:

    • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is a standard technique for protein separation.
    • Fixed-concentration gels have limitations in resolving proteins across a broad molecular weight range or those with similar molecular weights.

    Purpose of the Study:

    • To describe a protocol for pouring linear gradient polyacrylamide gels.
    • To highlight the advantages of linear gradient gels over fixed-concentration gels for protein separation.

    Main Methods:

    • Utilizing a gradient former to create gels with linearly increasing acrylamide concentration.
    • Pouring linear gradient polyacrylamide gels, commonly with a 4%-20% acrylamide range.

    Main Results:

    • Linear gradient gels allow for the separation of a broader range of protein molecular weight (M(r)) values.
    • Gradient gels increase the likelihood of resolving proteins with very similar M(r) values.

    Conclusions:

    • Linear gradient gels provide enhanced resolution and a wider separation range for proteins compared to traditional fixed-concentration gels.
    • The choice of acrylamide concentration range in gradient gels should be tailored to the specific proteins being analyzed.