Lab Expectations

This is part two of a two-part series on how to optimize your IHC protocols. Part one introduced the principles behind antigen retrival. Click here if you missed it... but, if you've got your tissue prepped and ready to go, we'll move on to the next steps in the staining protocol.

It’s Friday night and you could be out with your friends right now, but instead you’re tucked away in a dark little room filled with microscopes. Spending the evening in the lab seemed like a good choice at the time because you were certain this immunohistochemistry was going to reveal some small - but important - mystery of the universe to you. But now you’re sitting here, cursing the universe and everyone in it, because all you see when you stare down into the scope is some indistinct fuzziness. And did the controls work - meh - who’s to say? There’s no sugar coating it. It’s a fail.

Analysis of Immune Checkpoint Control Protein Co-expression in Breast and Ovarian Cancer Using Novel Rabbit Monoclonal Antibodies and Multiplex IHC

With an increasing number of biomarkers and, often, limited availability of biopsy material, there is a growing need for multiplexed assays for both research and clinical purposes. IHC based solutions are particularly attractive in the field of immuno-oncology, as maintaining spatial context within the tumor microenvironment provides meaningful and potentially actionable information.

Watch the video below.

Analysis of Immune Checkpoint Control Protein Co-expression in Breast and Ovarian Cancer Using Novel Rabbit Monoclonal Antibodies and Multiplex IHC

With an increasing number of biomarkers and, often, limited availability of biopsy material, there is a growing need for multiplexed assays for both research and clinical purposes. IHC based solutions are particularly attractive in the field of immuno-oncology, as maintaining spatial context within the tumor microenvironment provides meaningful and potentially actionable information.

Watch the video below.

This is part two of a two-part series on how to optimize your IHC protocols. Part one introduced the principles behind antigen retrival. Click here if you missed it...but, if you've got your tissue prepped and ready to go, we'll move on to the next steps in the staining protocol.

It’s Friday night - you’re tucked away in a dark, little room filled with microscopes.You could be out with your friends right now, but you begged off because you were certain this immunohistochemistry was going to reveal some small - but important - mystery of the universe to you.

Instead, you’re sitting here, cursing said universe and all its inhabitants, because all you see when you stare down the barrel of the scope is some indistinct fuzziness. And did the controls work - meh - who’s to say? There’s no sugar coating it – it’s a fail.

So, what’s next? If you’re like me, you’ll wing the slide into the trash with as much gusto as you can muster and head out to find your friends. Tomorrow, you’ll re-evaluate the experiment.

But where do you start? You probably know that a highly specific, high-affinity primary antibody is key to a successful IHC. But did you know that the companion reagents (i.e., buffers, etc.), which establish the pH and ionic strength of the system, are just as important? These reagents can influence the binding of the primary antibody to its epitope and dramatically affect the outcome of the assay.

To help you pick the best reagents for your assay (and make sure those Friday nights in the lab are worth the effort) we will spend the next several posts reviewing how companion reagents affect IHC. And, as an example we will describe our experience optimizing the protocol for one of our antibodies, PLK1 (208G4) Rabbit mAb.