Lab Expectations

With the arrival of the New Year, we all look back on 2017 and anticipate the coming year. CST scientists have been working hard to develop new antibodies and reagents to enable your next successful experiment in 2018!

In Roman mythology, the New Year and the month of January are associated with Janus, the god of transitions and doorways (1). Janus is best known for having two faces: one looking to the past, and one to the future. Janus also lends his name to the Janus kinase (Jak) family of nonreceptor tyrosine kinases.

You’re gathering data from all your experiments and preparing to present to your advisor and thesis committee at your annual progress report. You have an interesting hypothesis, and you have a validated antibody that recognizes your target protein on a western blot (WB). The molecular weight of the band is correct, and the expression of the target protein changes just the way you predicted it would. Now, you know — and you’d bet the house on it — when that powerpoint slide comes up, someone on your committee is going to ask about loading controls.

Early exploration of unmapped biological signaling pathways were carried out using radiolabeled phospho-imaging. The development of phospho-specific antibodies to detect and quantify protein phosphorylation made life easier for researchers (less ³²P waste to deal with), but the interpretation of data from these experiments comes with its own set of caveats. 

Part four of a series on immunofluorescence techniques. Check out previous posts on Validation, Experimental Controls, and Fixation/Permabilization. 

After your samples have been prepared, it's time to incubate them with a well-validated antibody, the workhorse of immunofluorescence. If you are a seasoned pro at IF experiments, you are probably used to checking the antibody datasheet (or web page) for the recommended dilution. But have you ever wondered where those recommendations come from?

The Zika virus (ZIKV) is related to flaviviruses that cause dengue and yellow fever, and is spread via mosquito (Ae. aegypti and Ae. albopictus) bites and secondarily via sexual transmission. ZIKV infection in adults is associated with mild dengue-like disease or Guillain-Barré syndrome, an autoimmune disorder of the peripheral nervous system. Most alarmingly, ZIKV is also transmitted across the placental barrier during pregnancy and can lead to the severe birth defect microencephaly (underdevelopment of the brain).

Part Three of a four-part series on Immunofluorescence. Check out The Importance of Validation and Experimental Controls.

The performance of an antibody is a crucial determinant in getting reliable immunofluorescence (IF) results. Equally important is the preparation of the biological sample - cells or tissue used in your experiments - before any antibodies are introduced. The fixation and permeabilization of your samples are key steps that can determine your experiment’s failure or success. The ideal fixative preserves a “life-like” snapshot while quickly stopping the degradative process of autolysis by crosslinking and inhibiting endogenous enzymes. This post provides examples of how different antibodies perform at their best using different protocols.

 Part two of a four-part series on Immunofluorescence. Check out our posts on Validation and Fixation and Permeabilization. 

After months of hard work, your research has honed in on a hypothesis you can test with immunofluorescence (IF). You've chosen antibodies and performed pilot IF experiments (see The Importance of Validation), and the localization of the protein appears reasonable. But how can you be sure the IF data you've acquired represents real biological phenomena? We present two examples of experimental controls in this post.

Part one of a four-part series on Immunofluorescence. Check out Experimental Controls and Fixation and Permeabilzation.

After months of hard work, your research has zeroed in on a hypothesis you can test with immunofluorescence (IF). But now you have to make a choice. How do you decide which antibody to use to get reliable IF results? How do you know if the images are accurately reporting the target's localization? We explore some considerations in this post.

The holiday season is traditionally marked by festive choral music, decorated cookies, and in recent years, ugly sweater contests. CST employees get to experience all three traditions at the same time at the company’s yearly Holiday Concert.