Total Protein Normalization - Advanced Tutorial
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- Опубликовано: 9 сен 2024
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Learn how to normalize fluorescence western blot signal to total protein using Image Lab.
In this video, we take an in-depth look at how to normalize your multiplex fluorescence western blots to total protein signal. We cover manual adjustment of lanes and bands, correcting for blot background, and how to export the data you need to publish these results.
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Hi. How can I measure the expression level of 3 proteins in the same blot?. I mean, After transfer, I first get the total protein image and once I´ve got the image in the software can I cut the membrane in 3 pieces to incubate each piece of membrane with a primary Ab, my secondary Ab, and then I either at the same time or separately get the image of three proteins (by chemiluminescence) and do the same as this tutorial?. or. it is impossible to cut the membrane and each protein has to be evaluated one by one?.
you better to do an immunoblot meaning that your secondary antibodies are conjugated to fluorescence molecules all antibodies are added at the same time too.