An Exploration of the Sigmoidal Curve – Math for the Rest of Us

An Exploration of the Sigmoidal Curve – Math for the Rest of Us

ELISA (Enzyme-Linked Immunosorbent Assay) is the heartbeat of many labs in the research world, owing to its simplicity and its ability to answer a very basic question: how much of protein/peptide/antibody is in my sample?  More specifically, it can be used to answer such questions as: How much IgG is in the serum after I…

Immuno-PCR:  A Highly Sensitive Method of Immunodetection

Immuno-PCR: A Highly Sensitive Method of Immunodetection

Researchers have relied on immunodetection techniques such as Western blotting, flow cytometry and Enzyme-Linked Immunosorbent Assay (ELISA) for years, but immuno-PCR is a relatively new method. By merging an ELISA with the Polymerase Chain Reaction (PCR), immuno-PCR provides extremely high levels of assay sensitivity. ELISA An ELISA is an assay in which a molecule is…

Ten Tips for Pipetting the 384-Well Plate

Ten Tips for Pipetting the 384-Well Plate

I was so excited to start using 384-well plates for my assays. With so many wells, these plates are useful for testing many conditions in parallel, as required in ELISAs, siRNA library screens, and drug treatment dilutions. However, I quickly learned that pipetting in these plates is more complicated than I thought. This article contains…

A Primer on Checking the Methylation State of the Genome

A Primer on Checking the Methylation State of the Genome

We all know that genes encode proteins that make up a living cell. However, the level and coordination of gene expression is really the key to the success of a living cell. One way eukaryotic cells (that’s us!) control protein expression is through addition of a methyl or hydroxymethyl group on the cytosine nucleotide. This…

A New Frontier in Protein Quantitation: AlphaLISA

A New Frontier in Protein Quantitation: AlphaLISA

If you ever worked in a biology or biochemistry laboratory, you probably already heard about ELISA. You may have even used it. But do you know what’s behind it? And how you can improve it? Let me guide you through the basics of ELISA, and introduce you to my favorite ELISA technique—AlphaLISA. First Things First… So,…

Are Proteins Adsorbing to Your Labware?

Are Proteins Adsorbing to Your Labware?

One of my favorite things about being a biochemist is to imagine everything at the molecular level—sometimes, in very corny ways. I envision the proteins I pipet and mix as dynamic characters in a molecular soap opera that intermingle with each other in complex ways. The biomolecular characters in my soap opera interact and react,…

Roadside Assistance: Fixing Your Broken-Down ELISA

Roadside Assistance: Fixing Your Broken-Down ELISA

The ELISA (enzyme-linked immunosorbent assay) is arguably one of the most important and versatile tools in the toolbox of molecular biologists, biochemists and diagnosticians across the world. Defined by its simplicity and speed, the assay is easy to learn and perform in as few as five steps. But with so few variables to manipulate, an…

Let me introduce you to ELISA…No, not the girl…The assay.

Let me introduce you to ELISA…No, not the girl…The assay.

An ELISA (Enzyme-Linked ImmunoSorbant Assay) is a popular assay that uses antibodies and color change to detect proteins, peptides, antibodies or biomolecules in complex mixtures. ELISAs are popular because they are reliable, specific, easy to use, and can easily be scaled up to process multiple samples simultaneously. How an ELISA is Done: In an ELISA,…

Tips for Choosing and Using a New Primary Antibody

Tips for Choosing and Using a New Primary Antibody

Finding a good primary antibody can often feel like playing Russian roulette.  Nothing is more disappointing than buying a $300 antibody that doesn’t work for your use. There are some steps you can take, however, to increase your likelihood of success. Scout out Other Labs Before you buy, ask if anyone around you or in…

5 Ways to Improve Your ELISAs

5 Ways to Improve Your ELISAs

ELISAs (enzyme-linked immunosorbent assays) are often used for detecting and quantifying substances such as peptides, proteins, antibodies and hormones in research and diagnostics. Today, a wide range of ELISA formats exist to suit your needs e.g. indirect ELISA, direct ELISA, competitive ELISA and sandwich ELISA. While it has become easier to perform ELISAs, thanks to…