Mar 20, 2024

Public workspaceImmunoblotting (Western) Protocol

DOI
dx.doi.org/10.17504/protocols.io.5qpvo3e3bv4o/v1
  • Scott Vermilyea1
  • 1University of Minnesota
Jane Balster
Open access
DOI: 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
Abstract
This protocol details the western blot technique.

Attachments
Western Blot and Lysate Prep Protocol.docx
Western Blot and Lys...
19KB
Immunoblotting (Western)
Immunoblotting (Western)
9h
Run protein lysates using desired Criterion™ TGX™ gels.
Transfer onto nitrocellulose membranes.
Detect protein of interest using appropriate primary antibodies at Temperature4 °C DurationOvernight , followed by horseradish peroxidase (HRP)-conjugated secondary antibodies for Duration01:00:00 at TemperatureRoom temperature .

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