Infrared Fluorescence Imaging: Making Western Blots More Quantitative
This webinar discusses the differences between fluorescent and chemiluminescent Western blot detection, multiplexed fluorescence detection. It also describes how you can easily transition from chemiluminescence, using ECL or other chemiluminescent substrates, to fluorescence detection. Examples of quantitative IR Western blots used in RNAi studies are given.
For more information, visit the quantitative Western blot application pages.
Date: December 12, 2009
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Speaker(s):
Amy Geschwender, Ph.D.
Principal Scientist
LI-COR Biosciences
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Advance video to...
- Introduction and Outline - 0:00
- IR Fluorescent Western Blotting - 3:56
- Limitations of Chemiluminescence - 6:23
- IR Fluorescent Western Blotting gives Direct Detection - 11:29
- IR Fluorescence is Different - 12:13
- Adjust the Image Display - 15:22
- Questions - 16:16
- Linear Range and Reference - 18:12
- Accurate Quantification with Reference - 22:14
- Multiplexing and Sensitivity - 24:01
- Lasers and the Odyssey Infrared Imaging Systems - 25:22
- Keys to High Sensitivity and Examples - 26:44
- IR Fluorescent Western Blotting is a Validated Technology - 31:19
- Questions - 34:09
- Quantifying with IR Fluorescent Westerns -36:29
- Analysis of Phosphorylation with Multiplex Westerns - 39:11
- RNAi Knockdown Validation - 41:00
- In-Gel Western Detection - 43:01
- Questions - 44:58
- Easy Transition from ECL Substrate - 47:31
- Chemi-IR Detection: A Bridging Technology - 48:27
- Blocking Buffers Are Important - 50:00
- Complete Solution for Quantitative Westerns - 52:35
- Questions - 56:22
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