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Western Blotting Reagents Roundup – Recent Publications

Azure offers products to support every step of your Western blotting workflow, from blocking to detection. Several recent publications highlight the use of Azure reagents in chemiluminescent and fluorescent Western blotting, in a variety of experimental systems and approaches. Below is a brief summary of four articles in which researchers incorporated Azure’s protein-free blocking buffer, […]

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Figure 1. AzureSpot Multiplex Image

Visualizing and Quantifying phosphoproteins via Western Blotting Part 2 of 2

Best practices for quantitation of unphosphorylated vs phosphorylated forms In part 1 we reviewed tips for using multiplexing to visualize phosphoproteins, and in part 2 we’re going to cover quantitation. Once you have reliable, repeated samples that fluoresce in non-overlapping bands and are linearly proportional to the signal, you can begin analysis. Here we will […]

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Visualizing and Quantifying phosphoproteins via Western Blotting Part 1 of 2

Western blotting is an important tool for researchers studying signal transduction and other processes where understanding the phosphorylation state of a protein is important for elucidating protein function. But some researchers are hesitant to use Western blotting to calculate the relative amount of phosphorylated to unphosphorylated protein. Here we discuss steps you can take to […]

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Loading controls and antibody validation for Western blot quantification

Today’s post is inspired by a conversation I had with a colleague when teaching them SDS-PAGE and Western blotting. Being well versed in qRT-PCR (R) and flow cytometry (R) they were particularly interested in how and why certain loading controls were used, and what other techniques were used to validate loading, changes in protein expression […]

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