The Ins and Outs of Protein Concentration – Semi-permeable Membranes

The Ins and Outs of Protein Concentration – Semi-permeable Membranes

This is the first of a three part series describing some of the most common methods for concentrating proteins. In later installments I’ll discuss using protein precipitation and chromatography to concentrate a protein. However, here I’ll detail the most popular approach – semi-permeable membranes, used for both dialysis and commercial protein concentrators. Structure of the…

So You Want to Work With Enzymes: What Is An Enzyme?

So You Want to Work With Enzymes: What Is An Enzyme?

Protein kinases and protein phosphatases phosphorylate and dephosphorylate a plethora of proteins. They are responsible for regulating the majority of cellular activities. Because of their importance, they can seem intimidating to tackle as a research project. At the end of the day however, kinases and phosphatases are- simply put- enzymes. Therefore, you can standard enzyme…

Basics of Protein Phosphorylation Part II: Tools of the Trade

Basics of Protein Phosphorylation Part II: Tools of the Trade

In the previous article in this series, we looked at the major players involved in protein phosphorylation: protein kinases, protein phosphatases, and target proteins. This time, we’ll glance over some of the tools that we can use to study various aspects of protein phosphorylation, focussing on a few I’ve personally come across. 1. Tools for…

Basics of Protein Phosphorylation Part I: Introducing the Candidates

Basics of Protein Phosphorylation Part I: Introducing the Candidates

The focus of my grad studies and postdoctoral research has been the analysis of proteins regulated by reversible protein phosphorylation. However, the number of unique facets in which protein phosphorylation can be studied is astounding, and is diverse as any niche of the biosciences. This article is the first in a series that will give…

Solved: Low Yields in Cell-Free Protein Synthesis

Solved: Low Yields in Cell-Free Protein Synthesis

In my last article, I introduced Cell-Free Protein Synthesis. Today I want to talk about a major bottleneck in in vitro cell-free protein expression; low yield. Most often, paying attention to the important factors such as template purity and design, transcription and translation inhibitors, and potassium and magnesium concentration will solve any problems with low…

An Intro to Cell-free Protein synthesis

An Intro to Cell-free Protein synthesis

Cell-free protein synthesis (aka In vitro translation) refers to protein production in vitro using lysates generated that provide the cellular machinery necessary for synthesis. The lysates can be of bacterial or eukaryotic origin. It is a useful alternative to in vivo synthesis for generating protein for the study of things like: protein:protein interactions (pull-down assays,…