Analytical Chemistry and Chromatography Techniques
How Separation Works in Column Chromatography Methods
Separation in column chromatography relies on differences. Molecules vary in size, charge, polarity, and solubility. We leverage these differences to distribute molecules between a stationary phase and a mobile phase. But because molecules are so different, it’s not possible to have a single method that works for all. In my previous article I discussed the…
Read More Fine-Tune Your MALDI-TOF to Produce Good-looking Mass Spectra
Mass Spec is all about getting the perfect peaks. Without a good peak assigning the correct mass is impossible and you cannot make accurate identifications. Make sure you know how to adjust your MALDI-TOF instrument parameters to achieve your perfect peak. In our previous posts ‘How does Mass Spec Work’ and ‘Imaging Mass Spectrometry: the…
Read More The Basics of Running a Chromatography Column
Column chromatography is a common technique used to separate individual compounds from a mixture. You can use column chromatography on both a small or large scale to isolate and purify material for use on a later experiment. If you are new to column chromatography or just want a refresher, then read on for a quick…
Read More Troubleshooting Thin Layer Chromatography: Some TLC for Your TLC
The whole TLC technique sounds easy to do, but it can be difficult and tricky during interpretation or give unexpected results, especially when working with biomolecules. For this reason, it is important to be familiar with troubleshooting thin layer chromatography. Some of the common problems faced during TLC and their solutions are listed below: Solvent…
Read More How to Choose Your MALDI (Soul) Matrix
gif by Kronin Okay not talking about that kind of matrix. But the kind of matrix that is crucial for MALDI (Matrix-Assisted Laser Desorption Ionization). I talked briefly about MALDI in my last article, Imaging Mass Spectrometry: The New(ish) Kid on the Block. Now I want to tell you more about MALDI, specifically the matrix.…
Read More How to Measure Molecular Weight with Gel Permeation Chromatography (GPC)
So you have a new polymer, unknown protein, or material combination but don’t know its molecular weight? An easy way to measure molecular weight of your sample is gel permeation chromatography (GPC). GPC is an analytical technique that separates molecules in polymers by size and provides the molecular weight distribution of a material. GPC is…
Read More Thin Layer Chromatography: The Basics
Like most other chromatographic techniques, thin layer chromatography (TLC) separates out individual compounds from a mixture depending upon the polarity of each compound. The solvent system travels up a silica plate by capillary action and passes over the sample that you spot onto the plate. As the solvent travels up, it moves the compounds present…
Read More The Many Uses of Thin Layer Chromatography
While working as biologists, we often come across mixtures of compounds, and the first question that strikes our minds is ‘what are the components in this mixture?’ One might think of using chemical assays to find the presence of specific compounds. But that sounds painful, doesn’t it? Well, the good news is that thin layer…
Read More Carrying You Through Gas Chromatography
Maybe you found a paper in which they used gas chromatography (GC) to analyze a sample that is similar to yours and now you are wondering if you should try it too. If you are hesitant because you know little about GC, then this article will help you with that. You may already know that…
Read More Cell Counting with a Hemocytometer: Easy as 1, 2, 3
Many biological applications such as microbiology, cell culture, blood work and many others that use cells require that we determine cell concentration for our experiment. Cell counting is rather straightforward and requires a counting chamber called a hemocytometer, a device invented by the 19th century French anatomist Louis-Charles Malassez to perform blood cell counts. A…
Read More What Can NMR Do For You – Part Three
Welcome to part three of “What Can NMR Do For You” This is an ongoing series in which we describe how you can use NMR experiments to characterize your protein of interest. In part one, we explained some critical points when doing one-dimensional (1D) NMR experiments on proteins and nucleic acids (e.g. sample preparation and…
Read More Getting the Most Out of Your Column: Optimizing Your HPLC Gradient
Let’s imagine the following scenario: You are researching a biosynthetic pathway in your favorite fungus. You know that this pathway produces a family of toxic compounds, and you want to see if you can block this pathway (or parts of it) with an antifungal drug. You have a control (no antifungal) and samples that have…
Read More How You Can Use HPLC In Your Research
If you’re an HPLC guru, then you probably think that everyone should be using HPLC. And you might have a point – HPLC is very powerful and has broad applications across many fields. But it isn’t the answer to every problem. HPLC (high-performance liquid chromatography) is used to separate mixtures of compounds based on their…
Read More The Seven Deadly Sins of HPLC
If you work with an HPLC, then you know the frustration of going to use the machine and finding it in disarray. If you’re new to using an HPLC, then the machine can be intimidating to use and you might not know the ins and outs of using it. Here’s an article that has a…
Read More Running You Through HPLC: How Does it Work?
Your advisor tells you that he wants you to use HPLC to analyze your compound. You know that you’ve heard of this technique before, but you can’t remember what HPLC stands for, let alone how to go about doing it! We’ve all been there, though. Fear not! In this article, we will remind you about…
Read More Running You Through HPLC: How Does it Work?
Your advisor tells you that he wants you to use HPLC to analyze your compound. You know you’ve heard of this technique before, but you can’t remember what HPLC stands for, let alone how to go about doing it! We’ve all been there, and I bet you wish you had paid more attention in that…
Read More Exploiting HPLC and UHPLC Selectivity With Rational Stationary Phase Design
In this seminar, we explore the power of selectivity using the resolution equation in HPLC and UHPLC. Using a variety of chromatographic data, we discuss how incorporating an aromatic building block in phase design has led to the unique bonded phases ACE® C18-AR and ACE® C18-PFP. These unique phases have been engineered based upon the popular…
Read More What Can NMR Do For You? — Part Two
Welcome to part two of “What Can NMR Do For You?”, a three-part series in which we see how you can use simple NMR experiments in your research. In part one, we went over some key points to keep in mind when doing NMR on proteins and DNA, such as sample preparation, and saw how…
Read More Recent Developments in Fused-Core Technology for the High Speed High Resolution Analysis of Biomolecules
HPLC separations of biomolecules have traditionally required much time in order to achieve the required resolution for complex samples. However, this has recently begun to change with the developments that have been made in Fused-Core particle technology. HPLC columns of high purity silica that are 2.7 µm in total particle size with a 0.5 µm…
Read More 5 Laboratory Sterilisation Methods
Effective sterilisation techniques are essential for working with isolated cell lines. For obvious reasons, you don’t want bugs from the environment growing in your nice culture medium, and equally, cultures must be sterilised before disposal. Unless you are using microbiology microscopes to view your cultures, you may be clueless about any unwanted microbe guests. Time…
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