Mar 21, 2024

Immunoblotting (Western) Protocol

DOI
https://dx.doi.org/10.17504/protocols.io.5qpvo3e3bv4o/v1
  • Scott Vermilyea1
  • 1University of Minnesota
  • Team Lee
Jane Balster
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DOI: https://dx.doi.org/10.17504/protocols.io.5qpvo3e3bv4o/v1
Protocol Citation: Scott Vermilyea 2024. Immunoblotting (Western) Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvo3e3bv4o/v1
License: This is an open access  protocol  distributed under the terms of the  Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 26, 2024
Last Modified: May 31, 2024
Protocol  Integer ID: 95764
Keywords: ASAPCRN, western blot technique, protocol this protocol detail, protocol detail, protocol
Abstract
This protocol details the western blot technique.

Attachments
Icon for Western Blot and Lysate Prep Protocol.docx
Western Blot and Lys...
19KB
Immunoblotting (Western)
9h
Run protein lysates using desired Criterion™ TGX™ gels.
Transfer onto nitrocellulose membranes.
Detect protein of interest using appropriate primary antibodies at 4 °C Overnight , followed by horseradish peroxidase (HRP)-conjugated secondary antibodies for 01:00:00 at Room temperature .

9h
To visualize, use chemiluminescent substrates and image using ImageQuant LAS 4010 detection system.
Perform densitometry on Western blot images using ImageQuant TL 8.1 software.