Optimize Chemiluminescent Westerns on the Odyssey® Fc Dual-Mode Imaging System

You do chemiluminescent Western blots – you do TONS of them. Sometimes, as I am sure you have discovered, you have ‘challenges’ – the darn thing just doesn’t look right!

What are some things that may happen when using chemiluminescence detection for Western blotting and how do you resolve them? The next series of blog posts here will offer up some tips on how to optimize chemiluminescent Westerns. These tips were designed for Westerns detected with a digital imager, like the Odyssey Fc Chemiluminescent and Near-infrared Fluorescent Imaging System. But many will also apply if you are (still) in the darkroom and using film. 🙂

High Background

Pools of excess substrate on the membrane can lead to areas of high background, as can adding more substrate to a membrane that has dried. Apply the substrate carefully and wick off any pools of substrate before imaging. Do not allow the membrane to dry.
pooled substrate

The image above is an example of high background due to pooled excess substrate. If your blot looks like the image below, then the membrane dried out and then someone tried to add more substrate.

dried blot

Optimizing Chemiluminescent Blots on the Odyssey Fc Imager

Introducing the NEW Odyssey® CLx Infrared Imaging System!

The Best Just Got Better!

Western blotting and the Odyssey CLx
Placing a Western blot on the New Odyssey CLx Imaging System

We at LI-COR are SUPER proud to introduce the new Odyssey CLx Infrared Imaging System. It’s the next generation Odyssey Imager – you know, the one that’s been the industry-leading quantitative Western blot technology for over 10 years!

The Odyssey CLx retains ALL of the application versatility and functionality of the Odyssey Classic. PLUS it now uses a great new software to get you and your lab up and running fast. . .so you can image all of your near-infrared fluorescent data and PUBLISH!

Image Studio™ Software is an extremely simple and easy-to-use imaging software. It is compatible with Odyssey® CLx and Odyssey Fc Imaging Systems.

  • Easy to use – training for new users is fast and simple
  • Intuitive, application-driven ribbon interface
  • Includes nine different types of analysis, including Western blots, DNA gel documentation, and small animal imaging