Techniques / Protein Expression & Analysis

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Why Do Enzymes Have Optimal Temperatures?

Every biologist is familiar with the profile of the rate of an enzymatic reaction versus temperature as shown in the figure. We know that enzymes from E. coli or warm-blooded animals tend to have an optimum...
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Article

Breakthroughs in Peptide Translocation: Cell Penetrating Peptides

Cell Penetrating Peptides (CPPs) are the Trojan Horse of cell biology – an innocuous peptide sequence with the special ability to carry virtually any cargo across the plasma membrane. If you have a special...
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Article

How to Make Your Own Luck in a Biochemistry Lab

During one of my first ever poster sessions, I was asked how I was able to make a particularly difficult-to-work-with protein soluble. My response was that I was just lucky. The venerable professor chuckled and...
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Article

How To Make Your Own ECL

ECL can be an expensive reagent in a lab, and what with it being involved in the final stage of a western blot, it’s something you don’t want to have to worry about too much. During my PhD, I was struggling...
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Article

Isoelectric Focusing for separation of proteins and peptides

SDS-PAGE is the standard technique used for separation of proteins in the lab, but that doesn’t meant that other techniques don’t have their place–one such technique is isoelectric focusing (IEF). IEF,...
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Article

The Practical Guide to Running a Perfect Homemade SDS-PAGE Gel

As biochemists, we routinely run SDS-PAGE to analyze our proteins. It can be a repetitive and tedious task to get a “presentable” gel for publication (especially when there are Western blots to follow)… but not if you know the tricks!
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Article

Split ubiquitin yeast two-hybrid

If you’ve read our article, An Overview of Yeast Two-Hybrid (Y2H) Screening, you’ll know that one major limitation of conventional Y2H is that your protein-protein interaction must occur in the nucleus for...
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Article

The Wet-chamber Method: How to use Less Antibody and Save Money

Want to save money on one of the most expensive steps of your Western blots? Then, use less antibody for significant cost savings!
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Article

Overview of In Planta Sub-Cellular Localization by Agroinfiltration

Agroinfiltration is a method for the transient expression of your protein of interest in a plant system. You can use it for the production of recombinant proteins or simply to determine the sub-cellular localization of your protein.
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How to Shut Off Background Lac Expression in LB

Here's a tip that you may find useful if you are expressing proteins in E.coli using a lac promoter-based expression system, e.g. pET, in LB medium (L-broth). Lac expression systems are typically induced in...
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