Posts Tagged ‘protein detection’
4 Fixatives for Histology and Cytometry. Perfect Your Preservation
Learn about four fixatives for histology, which one you should pick, and how. Plus, get some top tips for perfect sample preservation.
Read MoreGas Chromatography: What It Is, How It Works, and 5 Critical Components
Do you need to learn about gas chromatography? This article takes you through the basic principles and instrumentation. With illustrations!
Read MoreStrengths and Limitations of the Amazing NanoDrop™ Spectrophotometer
With the NanoDrop spectrophotometer, quantifying a DNA, RNA, or protein sample concentration is as easy. Here’s everything you need to know about the strengths and limitations of this handy spec.
Read MoreProtein Staining Methods: An Overview of 3 of the Best
There are several great protein staining methods, but how do you pick the one that’s appropriate to your intended application? Read on to find out.
Read MoreThe 3 Most Common Flow Cytometry Fallacies
Flow cytometry is fast evolving from a method only revered by immunologists, to one used by nearly every biological specialty. It’s pretty much my favorite tool. Unfortunately, as with most lab techniques, much of flow cytometry is taught on the job without a lot of standards. And too often bad habits are passed along like…
Read MoreHow to Store Your Reagents, so They ‘Do Exactly What It Says on the Tin’
Your reagents should do ‘Exactly what they say on the tin.’ This only happens though if you look after them in the way the manufacturer states on their data sheets. We have all been guilty of using reagents past their expiration date. Usually we can get away with it, but there are a few things…
Read MoreFluorescent Western Blotting: Lowdown and Advantages
In this article, you will be introduced to the world of fluorescent western blotting. Firstly, we will compare fluorescent and chemiluminescent western blotting. Then, we will learn how infrared fluorescent western blotting can give you truly quantitative and reproducible results. Lastly, we’ll look at the many advantages of fluorescent western blotting, including the possibility to multiplex. Importantly,…
Read MoreHow to Scrutinize Your Glycosylated Proteins Without Using Glycosidases
You might have come across protein glycosylation before. Somewhere in the recesses of your memory you might even recall reading something about the protein you’re studying being glycosylated, but what does this mean and how do you analyze it? Glycosylated proteins are molecules decorated with sugar groups as they pass through the ER and Golgi…
Read MoreMultiplex Cytometric Bead Array: The ABCs of CBAs
Multi-parameter data acquisition is key to the modern era of science research. I, for one, wish every single experiment that I design would give me the maximum amount of information. For example, in cell biology and immunology, we want to capture as much information (be it cytokines/hormones/chemokines) as possible about a given cell population. Of…
Read MoreFour Tips for Working with Human Clinical Samples
While using human clinical samples in your research can provide robust and heterogeneous results applicable to larger portions of the population, working with these samples presents its own set of challenges. Here are some tricks I have learned to help isolate and grow your cells of interest while eliminating stromal, blood, or other undesired contaminants.…
Read MoreLighting the Way: Understanding Flow Cytometry Fluorophores
As science is becoming more interdisciplinary, the tools we use to answer questions are also crossing party lines. Case in point: flow cytometry. Once a tool only used by “real” immunologists, flow cytometry is fast becoming a method by which numerous questions can be answered, from the length of a cell’s telomeres, to the state…
Read MoreSDS-PAGE Tips and Tricks to Save You Time
Speed up your SDS-PAGE with our time-saving tips and tricks!
Read MoreHow to Cherry Pick Your Primary Antibody
How do you pick which antibody you should use in your assay? If you’re starting a new assay and need an antibody for the job, then selecting a new antibody from the plethora available could be high up on your to-do list.
Read MoreA Beginner’s Guide to Tag-Removing Proteases
Every biochemist is familiar with proteases. More often than not, proteases cause a lot of anxiety. To this end, a lot of research has been done in developing techniques to prevent the activity of proteases. But some of these proteases can be the good guys too! For example, you can use them to separate your…
Read MoreSonication – 7 Tips for Mastering the Art
Sonication is mostly used during preparation of protein extracts to help break apart the cell. Although most lysis buffers have buckets of detergent that lyse cell membranes, sonication just gives an extra hand in breaking everything apart. Sonication also breaks up, or shears, DNA in a sample—preventing if from interfering with further sample preparation. Have…
Read MorePolymers as Secondary Antibodies for Immunohistochemistry
Do you use biotinylated secondary antibodies in your immunohistochemistry? You could use polymers instead. They are a great time-saving reagent.
Read MoreCell Cycle Analysis by Flow: DNA Stains and Beyond
While you can observe mitotic cell cycle progression using immunofluorescence, flow cytometry is a great tool to delineate details that aren’t apparent by chromosomal morphology alone. DNA stains are a great way to get a general idea of what your cells are up to. There are also a number of other stains you can use…
Read MoreCommon Mass Spectrometry Contaminants: I Messed It Up So You Don’t Have To!
Through many trials, and lots of error, I learned that there are many considerations for mass spectrometry that might not be obvious to you as a molecular biologist. Common contaminants, even in small quantities, can mask important peaks in your mass spec data and have a huge impact on the final results.
Read MoreThe Messy World of Human Tissue Research
In neuroscience and other biomedical research fields, animal models allow us to answer critical but otherwise impossible questions. Despite the value of these models, however, sometimes nothing can replace the real deal—human tissue. The limitations of human tissue research stem from the variability between subjects and the risk for covariates that influence your results. Which…
Read MoreHow to Use Proteases to Purposefully Digest Proteins
In this article I will not talk about ‘wild’ proteases, which destroy cellular proteins in your lysates like wolves destroy sheep. Instead, I’ll be talking about the shepherd dog proteases—purified, tame and useful to digest proteins your research. In Protein Research and Crystallization Several programs can predict your protein domains. However, we wet biologists know…
Read MoreHow to Destroy your Flow Cytometry Data in 3 Easy Steps: Snap, Crackle, and Pop
While many scientists are methodical and precise, some of us like to live on the edge. Read a protocol all the way through? No thanks, I’ll take my chances and guess what concentration of HCl I should use. Label my tubes with the correct content? Puh-lease – it’s much more exciting deducing which is which…
Read MoreA Simple Method for Measuring Intracellular Fluorescence
Fortunately for microscopy users, measuring intracellular fluorescence has been made relatively simple through an ImageJ plugin called the Cell Magic Wand. For those of you unfamiliar with ImageJ, it’s a popular image processing program that runs on Mac, Windows, and Linux. How to use ImageJ for measuring intracellular fluorescence First of all, to begin measuring…
Read MoreTop 5 Tricks for Using FlowJo
Are you planning to do cellular immunology research? Then chances are you will be introduced to the flow cytometer – “a modern immunologist’s best friend.” This modern magic box is a highly versatile machine packed with cutting-edge fluidics and photonics (lasers). Combined with the monoclonal antibodies conjugated to fluorochromes capable of emitting light signals from a…
Read MoreTroubleshooting 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 MoreIsolating Proteins? Use Histidine and Transition to Metals
To isolate your protein, you are going to need to tag it with something that will allow you to fish it out of the bacterial protein soup. This is where transition metals can help you out!
Read MoreMass(ively powerful) Cytometry
Mass Cytometry is a relatively new technology which has recently featured in many high-impact journals. You may have read about instruments including the CyTOF, CyTOF2, and more recently, the Helios. With these instruments becoming more widespread, you might find yourself asking, what is mass cytometry, and what can it do for you? The Basis: Conventional…
Read MoreThe Practical Guide to Running a Perfect Homemade SDS-PAGE Gel
As biochemists, we routinely run SDS-PAGE to analyze our proteins. Imagine the time and effort you are going to save when you can run every gel to perfection.
Read MoreMultifocal Structured Illumination Microscopy: The Fast Food of Super-Resolution Techniques
While most of us have heard of super resolution microscopy, many of you may not have heard of MSIM, or Multifocal Structured Illumination Microscopy. This under-the-radar imaging technique is relatively quick, cheap (by comparison) and will allow you to get a lot of data, fast. So What is MSIM Anyway? MSIM, as I mentioned earlier,…
Read MoreGetting Started with Raman Spectroscopy: What You Need to Know
Are you an assiduous biologist who prefers label-free imaging methods for biological samples analysis? Raman spectroscopy offers you a wonderland of imaging technique with unlimited benefits. To start with, Raman Spectroscopy is a spectroscopic technique based on inelastic scattering of monochromatic light usually from a laser in the visible or near infra-red part of electromagnetic…
Read MoreWhere are My Bands? Troubleshooting a Signal-less Western
Western blotting uses electrophoresis and antibody-epitope affinity to give a semi-quantitative and (theoretically) clear measure of protein abundance. It’s a long procedure, filled with many steps—and even more room for error. Learning to troubleshoot certain problems is incredibly important for continued success with this technique. So what do you do when your final imaged product…
Read MoreTips for Taking Immunofluorescent Images for Your Next Paper
Taking publication quality immunofluorescent images of can be a very time intensive, and frustrating process with hours spent capturing, processing, and putting the images into final figure format. And, if you aren’t careful, you can do a lot of work only to realize later that you need to re-image something for one reason or another.…
Read MoreHow to See the Cell Cycle Through Your Microscope
Even in the most basic applications, fluorescence microscopy can be a very powerful technique. Simply put, the ability to actually see the biology you are interested in cannot be matched in directness. Often, the aim of fluorescence microscopy is to observe the effect of an experimental manipulation. Ultimately, you would like to know that the…
Read MoreThe 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!
Read MoreIntroducing Aptamers for Super Resolution Microscopy
What do you use if antibodies are too large for super resolution microscopy? Aptamers. These are small affinity reagents (~ 2 nm!) that interact with their target in the same way as an antibody, but without the hefty backbone attached.
Read MoreThe Key to Unlocking DNA from FFPE Tissues
Formalin fixed paraffin embedded (FFPE) tissues are valuable samples that typically come from human specimens collected for examination of the histology of biopsies for the detection of cancer. But each sample contains much more information just waiting to be unlocked. Despite the tiny sample size, DNA can be extracted from the tissue sections and used…
Read MoreHow Sweet is Your Protein: Using Enzymes to Study Glycosylation
Most eukaryotic proteins exist as several isoforms, differing in posttranslational modifications, which allows them to perform slightly different functions or the same function under slightly different conditions. A common posttranslational modification of proteins is glycosylation.
Read MoreImmunoscience or Immunoalchemy?
First of all let me say the technique of labeling tissues (immunohistochemistry, IHC), and cells (immunocytochemistry, ICC) is indeed immunoscience NOT alchemy, though at times it may certainly seem like alchemy! But to scientists inexperienced in this technique, who typically see the results of IHC/ICC experiments in the form of pretty pictures, it can certainly…
Read MoreNative Versus Denaturing Gels
We’re already gone through the basics of how gel electrophoresis work, compared common gel types like agarose and polyacrylamide and even explored some alternatives. Now let’s look at the native versus denaturing gels. You’ll be a speGEList in no time! Denaturing Gels We’ll start with this one, as it’s very self-explanatory. Denaturing gels are exactly…
Read MoreRemote Cytometry: Help from beyond!
The idea of accessing one computer from another is long established. Unfortunately, we often have visions of hackers sneaking in and stealing our data when we have most to lose. However, this type of technology can aid us in a lot of applications and to those of us who work in cytometry the benefits are (somewhat) clear. No More ‘Fail’ Moments Many researchers know the dread of…
Read MoreHow to Make Lipid Bilayers
Supported lipid bilayers are a very useful tool in many fields of cell and membrane biology. But how easy is it to make them? Bilayers can be made quite reproducibly, once you have found a reliable protocol! However, it can take some time to optimize your technique, so to increase your chances, make sure you…
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