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Quantitative Western Blotting: How and why you should validate your antibodies

Validating your antibodies, i.e.  confirming that an antibody recognizes your protein of interest with low cross-reactivity to other targets, is critical for ensuring consistent, reproducible results. Validation is frequently required before submission to a journal, but not everyone takes the time to validate. Or many of us assume that because we validated the antibody for […]

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New method effectively stains apoptotic retinal cells without requiring intraocular injections

Azure Biosystems is excited to announce the first published study that uses the Sapphire Biomolecular Imager, and we look forward to the role that Sapphire will play in future studies. What’s the problem, scientifically speaking? Labeling neurons is critical for the diagnosis, prognosis, and treatment of retinal degenerative diseases. A common approach is to use […]

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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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