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The evolution of the ELISA and other immunoassays

If you’re reading this blog there’s a high chance that you use antibodies in your work, in some form of immunoassay. However, many of us rarely stop and think about where these antibodies come from and how their development has been fundamental for the evolution of life sciences. A brief history of the antibody Antibodies […]

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Linear range of detection and what it means for your quantifications

When quantifying Western blots many people assume it’s ok to simply image your protein of interest, strip, re-probe for their loading control of choice and then use this to normalise, without considering the linear range of detection. However, there are several pitfalls associated with this, and more and more journals are now asking for a […]

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Which protein assay is best for you?

Have you ever spent ages correctly quantifying your protein samples, cast the perfect gel, efficiently transferred, probed and eventually come to the visualisation stage only to be presented with a loading control like this: Well before you throwing your blot out with the bathwater and repeating the experiment you should maybe consider your choice of […]

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Loading control example

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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In Cell Western
Protein Assays
Western Blotting