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Related Concept Videos

Western Blotting01:15

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
Types of Membrane Protrusions01:28

Types of Membrane Protrusions

The protrusion of the cell surface is an initial step for several cellular processes, including cell migration, phagocytosis, and neurite outgrowth. These membrane protrusions are a result of cytoskeletal rearrangement. The most  widely observed cell protrusions include lamellipodia, pseudopodia, filopodia, microvilli, invadopodia, and podosomes. These protrusions can be of two types — static or dynamic.
The microvilli, an example of stable protrusions, are finger-like projections with a...
Membrane Domains01:18

Membrane Domains

The membrane domains concentrate specific lipids and proteins at one place within the membrane, which helps in cell signaling, adhesion, and other critical cellular processes. These domains can differ in size, composition, function, and lifespan.
Protein Domains
The membrane comprises a group of distinct proteins responsible for carrying out a cell's specific function. For example, the plasma membrane of the human sperm, or a single germ cell, contains a unique set of proteins in the anterior...
What are Membranes?01:24

What are Membranes?

A cell's plasma membrane demarcates the cell's borders and determines the nature of its interaction with the environment. Cells exclude certain substances, take in others, and excrete some others in controlled quantities. The plasma membrane must be flexible to allow certain cells, such as red and white blood cells, to change their shape while passing through narrow capillaries. These are the more obvious plasma membrane functions. In addition, the plasma membrane's surface carries markers that...
What are Membranes?01:54

What are Membranes?

A key characteristic of life is the ability to separate the external environment from the internal space. To do this, cells have evolved semi-permeable membranes that regulate the passage of biological molecules. Additionally, the cell membrane defines a cell’s shape and interactions with the external environment. Eukaryotic cell membranes also serve to compartmentalize the internal space into organelles, including the endomembrane structures of the nucleus, endoplasmic reticulum and Golgi...
What are Membranes?01:24

What are Membranes?

A cell's plasma membrane demarcates the cell's borders and determines the nature of its interaction with the environment. Cells exclude certain substances, take in others, and excrete some others in controlled quantities. The plasma membrane must be flexible to allow certain cells, such as red and white blood cells, to change their shape while passing through narrow capillaries. These are the more obvious plasma membrane functions. In addition, the plasma membrane's surface carries markers that...

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Related Experiment Video

Updated: Jun 4, 2026

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins
11:11

Determination of Plasma Membrane Partitioning for Peripherally-associated Proteins

Published on: June 15, 2018

Far Western: probing membranes.

Margret B Einarson, Elena N Pugacheva, Jason R Orlinick

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary
    This summary is machine-generated.

    The far-Western blot assay uses GST fusion proteins to detect protein interactions on membranes, offering an alternative to traditional Western blotting. This method includes a renaturation step to improve protein interactions after transfer.

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    Published on: August 22, 2007

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Protein Interaction Analysis

    Background:

    • Western blotting detects proteins using antibodies.
    • Far-Western blotting (overlay assay) uses recombinant GST fusion proteins to detect protein interactions on membranes.

    Purpose of the Study:

    • To describe a modified far-Western blot protocol.
    • To enable detection of protein interactions that may be hindered by improper protein folding after membrane transfer.

    Main Methods:

    • Proteins are transferred to a membrane, washed, blocked, and incubated with a labeled or unlabeled GST fusion probe protein.
    • Detection can be radioactive (using 32P-labeled probe) or non-radioactive (using GST antibodies).
    • A denaturation and slow renaturation step is included to facilitate protein interactions.

    Main Results:

    • The protocol allows for the detection of target proteins interacting with GST fusion proteins on a membrane.
    • The renaturation step can improve the ability of transferred proteins to interact.

    Conclusions:

    • Far-Western blotting is a valuable technique for studying protein-protein interactions.
    • The described protocol enhances the assay's effectiveness by addressing protein folding issues.
    • Careful controls are necessary to distinguish specific interactions from non-specific binding and antibody cross-reactivity.