Author: Promega

Reaching Out for Lab Research Experience

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Today’s guest blog is written by Melissa Martin, a global marketing intern with Promega this summer. She will be a senior this fall at the University of Wisconsin-Madison where she is double majoring in zoology and life sciences communication, with a certificate in environmental studies.

Congrats! You are attending a university and pursuing a challenging, yet rewarding, undergraduate science degree. Getting to this moment probably included lots of late nights spent studying or worrying while applying to your dream college. However, now that you are here you will find that classes provide a lot of information. You can even take your education one step further by getting hands-on experience in a research lab.

Working in a lab is not only about making your resume look good. It offers a real-world experience that directly enhances your learning experience and can even guide your future. For example, your experiences in the lab can teach you basic skills (pipetting, determining concentrations, performing titrations, etc.) that will be useful in a variety of science professions.

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Immune Checkpoint Bioassays Strengthen Cancer Research in the Development of New Therapies

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This post was written by guest blogger, Nicole Werner, Product Management Support at Promega GmbH.

“You have cancer.” – a statement that fundamentally changes life in a second. After the first shock, the insight often arises: “If only I had stopped smoking sooner!”

Lung cancer, while not the leading cause of death worldwide, is the leading preventable cause of death in developed countries. According to the WHO, eight million people die each year as a result of smoking, including one million as a result of passive smoking [1]. Currently, 80% of those affected die within the next 13 months after diagnosis [1]. New therapeutic approaches, such as treatment with immune checkpoint inhibitors, bring hope.

Promega supports research in this area with the high-precision tools needed to develop this new form of therapy.

Artistic 3D rendering of Immune checkpoint signaling. Immune checkpoint bioassays enable researcher to characterize therapeutic antibodies trageting these pathways.
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RIPK1: Promising Drug Target of Chronic Inflammatory Diseases

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Today’s post is written by Michael Curtin, Senior Product Manager, Reporters and Signaling.

Inflammation is a defense mechanism that the body employs in which the immune system recognizes and removes harmful and foreign stimuli and begins the healing process. Inflammation can be either acute or chronic. Chronic inflammation is also referred to as slow, long-term inflammation and can last for prolonged periods (several months to years); chronic inflammation is caused by immune dysregulation. This typically takes the form of the body’s inability to resolve inflammation resulting from overproduction of inflammatory cytokines and chemokines, as well as danger-associated molecular patterns (DAMPs) released from dying cells (2). Tumor Necrosis Factor (TNF) is the primary cytokine involved in many common inflammatory diseases and is where many therapies targeting inflammation are focused.

Signaling of kinases like RIPK1 can be studied using the NanoBRET target engagement assays

Recent research that RIP kinases (RIPK1 and RIPK3) are important regulators of innate immunity via their key roles in cell death signaling during cellular stress and following exposure to inflammatory and infectious stimuli. RIPK1 has an important scaffolding role in pro-inflammatory signaling where it interacts with TRADD, TRAF1 TRAF2, and TRAF3 and TRADD can act as an adaptor protein to recruit RIPK1 to the TNFR1 complex in a TNF-dependent process. RIPK1 plays a kinase activity-dependent role in both apoptotic and necroptotic cell death. A review article by Speir et al. (1) discusses the role of RIP kinases in chronic inflammation and the potential of RIPK1 inhibitors as a new therapeutic approach for the treatment of chronic inflammation. RIPK1 or Receptor Interacting Protein Kinase 1 is a serine/threonine kinase that was originally identified as interacting with the cytoplasmic domain of FAS. Promega offers several reagents that make studying RIPK1 easier- these include our RIPK1 Kinase Enzyme Systems which includes RIPK1 (Human, recombinant; amino acids 1-327), myelin basic protein (MBP) substrate, reaction buffer, MnCl2, and DTT and is optimized for use with our ADP-Glo Kinase Assay.

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Overcoming Challenges to Detect Apoptosis in 3D Cell Structures

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This blog is written by guest author, Maggie Bach, Sr. Product Manager, Promega Corporation.

Researchers are increasingly relying on cells grown in three-dimensional (3D) structures to help answer their research questions. Monolayer, or 2D cell culture, was the go-to cell culture method for the past century. Now, the need to better represent in vivo conditions is driving the adoption of 3D cell culture models. Cells grown in 3D structures better mimic tissue-like structures, better exhibit differentiated cellular functions, and better predict in vivo responses to drug treatment.

Switching to 3D cell culture models comes with challenges. Methods to interrogate these models need to be adaptable and reliable for the many types of 3D models. Some of the most popular 3D models include spheroids grown in ultra-low attachment plates, and cells grown in an extracellular matrix, such as Matrigel® from Corning. Even more complex models include medium flow over the cells in microfluidic or organ-on-a-chip devices. Will an assay originally developed for cells grown in monolayer perform consistently with various 3D models? How is measuring a cellular marker different when cells are grown in 3D models compared to monolayer growth?

Close up of cells in 3D culture apparatus. 3D Cell Structures Provide Challenges for Measuring Markers of Cellular Activitiy
3D Cell Structures Provide Challenges for Measuring Markers of Cellular Activitiy
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Bringing Cutting Edge Technologies to Academic Researchers Through the Academic Access Program

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This post was written by guest blogger Iain Ronald, Director Academic/Government Market Segment at Promega.

My back story is similar to most of you reading this blog, high school education, undergraduate degree then onto a postgraduate degree. However, over 25 years ago during my undergraduate study, I was fortunate enough to work in the lab of Professor Ray Waters studying DNA damage in S. cerevisiae as a model organism and at the time PCR was cutting-edge technology and the PCR license was in full effect. However, there was one company that was fighting the good fight to democratize PCR for the good of the scientific community, Promega.

I became enamored with Promega then, and the next steps in my career were taken with a view to working at this company who, for all intents and purposes, seemed to really care about the progression of science beyond self-aggrandizement.

Now that I am working at Promega in a position where I can bring benefit to our academic community, I have pondered what I can do to equal the disruptive attitude I observed in this company all those years ago when they were fighting the then “big tech” for the enablement of the scientific community. 

Reporter bioassays are one of hte many offerings of the academic access program.
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How Promega Helped Our Lab Scale Up Drug Discovery for Bloodborne Pathogens

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This blog was written by Sebastien Smick, Research Technician in Dr. Jacquin Niles’ laboratory at Massachusetts Institute of Technology (MIT)

Our lab is heavily focused on the basic biology and drug discovery of the human bloodborne pathogen Plasmodium falciparum, which causes malaria. We use the CRISPR/Cas9 system, paired with a TetR protein fused to a native translational repressor alongside a Renilla luciferase reporter gene, to conditionally knock down genes of interest to create modified parasites. We can then test all kinds of compounds as potential drug scaffolds against these gene-edited parasites. Our most recent endeavor, one made possible by Promega, is a medium-low throughput robotic screening pipeline which compares conditionally-activated or-repressed parasites against our dose-response drug libraries in a 384-well format. This process has been developed over the past few years and is a major upgrade for our lab in terms of data production. Our researchers are working very hard to generate new modified parasites to test. Our robots and plate readers rarely get a day’s rest!

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Drug Repurposing Screens: Redeploying Old Dogs for New Tricks

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This blog was written by guest author, Amy Landreman, PhD.

Drug repurposing, identifying new uses for approved or investigational drugs, is an attractive strategy when looking for new disease treatments. Because the compounds have already gone through some level of pre-clinical optimization and safety testing, this approach can reduce risk, reduce cost, and speed up the timeline for further drug development. An additional benefit of this approach is that it can result in new biological insights or a better understanding of disease mechanisms since these compounds usually already have some level of mechanistic characterization. Indeed, there are now a number of compound collections openly available specifically for the purpose of facilitating drug repurposing efforts. For example, the ReFRAME (Repurposing, Focused Rescue, and Accelerated Medchem) library is a collection of 12,000 compounds developed by Scripps Research Center and has been screened to identify novel candidate therapeutics for Cryptosporidium infection (1). The Broad Institute also offers a drug repurposing hub that contains an annotated collection of over 7,000 compounds.

Drug repurposing libraries, although often smaller than novel compound small molecule libraries, are designed for implementation into high-throughput screening workflows in order to efficiently triage compounds for the desired result. Effective compound screens require assays that can be scaled to 384 or 1536-well microplate formats and implemented in batch or continuous processing workflows. The firefly luciferase reaction has been leveraged to create many assays that are well-suited to these types of high-throughput screening approaches. In particular, the generation of “Glow” assays that have stable luminescent signals and homogenous assay design is a good fit. The signal stability allows for multi-plate processing and because the reagent is added directly to cells in culture, pre-processing steps are eliminated allowing for automated workflows. Assay reagents such as the CellTiter-Glo® Cell Viability Assay and the ADP-Glo™ Kinase Assay are commonly used in screening efforts including those done with repurposing libraries.  In addition, there are several firefly luciferase reporter assay reagents such as Steady-Glo® and Bright-Glo™ Luciferase Assays that have been optimized for high-throughput detection of firefly luciferase activity making them well-suited to repurposing screens.

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The Power of Vulnerability

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Today’s blog is written by Malynn Utzinger, Director of Integrative Practices, and Tim Weitzel, ESI Architect.

If we want to reignite innovation and passion, we must rehumanize work.

-Silicon Valley CEO of Several Start-ups

If we want to rehumanize work, we need to be more human in the workplace.

-Promega’s ESI Bootcamp

Vulnerability is the birthplace of intimacy, trust connection, creativity, innovation. For leaders, it is the birthplace of trusted influence. But it is not permission to overshare.

-Brené Brown

Myths of Vulnerability

It’s important that we start off by making a few things about vulnerability crystal clear:  being vulnerable is not about over-sharing, being emotional—or worse, gushy. It is not about sacrificing necessary boundaries or letting go of all discernment when speaking. Vulnerability, as we intend it, is about being real with others. It is about being clear and honest enough within yourself that you can use courage and clarity to state a need or a perspective. Quite the opposite of requiring tears or grand displays of emotion, vulnerability can be expressed with utter command of one’s emotions, so that the clarity and authenticity of the message is what remains.

Vulnerability is also knowing that you cannot know everything or do your work perfectly or even to your full satisfaction sometimes, and it is having this same understanding and acceptance for others. It is being able to speak to that honestly so that we can build sustainable bridges between ourselves and others. We call this speaking our truths–with discernment.

Finally, vulnerability is knowing that while we must give our best efforts where and whenever we can, we must also know what we can’t control.  In most cases, what we cannot control is outcomes.  Therefore, vulnerability is embracing the uncertainty in how things will go in our relationships and in our work if we risk emotional exposure.  We cannot always know how others will hear what we share, but we can learn to take that risk and speak in service to a common goal.  For example, we might decide to share that the reason we are being so obsessive or insistent on a process is because of a past failure (perceived or real) that we still carry with us.  Even though we cannot control what others will think of our story, we trust that the sharing may help them share a need of their own or to hear our own need differently, so that we can all work together.  This is true in every relationship of our lives, where we learn to share something true for the sake of allowing another human being to know us as we are. 

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Reflections: King’s College London iGEM 2020, Renervate and Future Prospects

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Today’s guest blog about the 2020 virtual iGEM Giant Jamboree is written by Abigail Conner, Co-Team Leader of iGEM King’s College London (KCL).

In October 2019, I returned to London from Boston feeling elated after an unforgettable week at the Giant Jamboree. My team, Capacity, had just won a Silver Medal. I had the privilege of presenting in front of the judges about our work. The Giant Jamboree presented me with a vision of where Synthetic Biology will take us and its potential to radically transform our society for the better. Words cannot describe the deep sense of pride I felt to be a part of this community. For the first time, I felt truly empowered as a young scientist and was hugely inspired by the brilliance of my peers. As a result, I was beyond happy to assume the role of Team Leader of KCL’s 2020 team.

Almost immediately after touching down in the United Kingdom, I began to plan our project. Throughout the recruitment process and setting up applications, Stephanie Avraamides—the Head of Human Practices in Capacity—joined me in leading the team. As Co-Team Leaders, we would establish Renervate, a team of 19 undergraduate students from various STEM backgrounds, from Nutrition to Biomedical Engineering. Although we were fortunate to have met up in person several times before March, the onset of the COVID-19 pandemic scattered us across the world. Our team members represent sixteen different countries, meaning we had to navigate a range of time zones when working virtually. Despite this, we adapted to the virtual setting and worked tirelessly to develop Renervate. Come November, we would be rewarded for our endurance and commitment. I am thrilled to say that Renervate won a Gold Medal, Best Therapeutics Project, and nominations for Best Model and Best Supporting Entrepreneurship at last year’s Virtual Giant Jamboree.

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Technical Manuals: A New Look

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This blog post was cowritten by Sara Klink and Kari Kenefick.

Promega technical manuals have a new look! But never fear, our manuals still contain the protocol instructions for correctly using Promega products and include data, product and component storage information that you need to be successful at the bench. The cover art on our manuals now incorporates the use of imagery created by David Goodsell, which you can also find on our product boxes and at www.promega.com. The new cover image is being applied as we create new technical manuals or revise existing documents. Below are the old (left) and new (right) covers to compare:

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