The Protein Man's Blog | A Discussion of Protein Research

How to Prepare Samples for Western Blot Analysis

Posted by The Protein Man on Jan 21, 2020 2:30:00 PM

As a common technique in molecular and cell biology, Western blot analysis allows you to detect specific proteins in a tissue homogenate or extract. However, if you want to ensure the success of your Western blot experiment, you need to learn how to properly prepare samples.

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Topics: Western Blotting

Using Western Transfer for Protein Detection: Some Commonly Used Methods

Posted by The Protein Man on Jul 29, 2019 1:41:42 PM

Question:

How to detect protein using western blotting?  

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Topics: Western Blotting

Optimize Antigen and Antibody Concentration for Western Blots

Posted by The Protein Man on Jul 18, 2017 2:30:00 PM

Introduction

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Topics: Western Blotting

Western Blot Blocking: Tips and Tricks for Blocking Agents

Posted by The Protein Man on May 30, 2017 2:31:00 PM

Good Western blot blocking methods are crucial in order to achieve clean and reliable results. Protein samples are first ran on a polyacrylamide gel to separate proteins by size. The proteins are then transferred to a nitrocellulose or PVDF membrane. A blocking agent is then used to prevent primary and secondary antibodies from binding to the membrane non-specifically (areas without the protein of interest). Proper blocking methods cover the membrane surface without interfering with bound protein; thus preventing non-specific antibody binding and non-specific signal (noise or background) due to the membrane’s high affinity for proteins.

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Topics: Western Blotting

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