Bottom-up Proteomics: Need Help?

Posted on by Kassidy Steyer

The use of mass spectrometry for the characterization of individual or complex protein samples continues to be one of the fastest growing fields in the life science market.

Bottom-up proteomics is the traditional approach to address these questions. Optimization of each the individual steps (e.g. sample prep, digestion and instrument performance) is critical to the overall success of the entire experiment.

To address issues that may arise in your experimental design, Promega has developed unique tools and complementary webinars to help you along the way.

Here you can find a summary of individual webinars for the following topics:

Continue reading “Bottom-up Proteomics: Need Help?”

Here Comes the Sun: How to Protect Yourself and the Coral Reefs

Posted on by Kassidy Steyer

Sunscreen usage is increasing, with more people using SPF to prevent the very real threats of skin cancer and early signs of aging. While slathering on the sunscreen is unarguably important to protect your skin from the sun, new concerns arise linking sunscreen chemicals to coral reef bleaching, as an estimated “14,000 tons of sunscreen is believed to be deposited in the oceans annually.”

Coral reefs are the most productive marine ecosystem known. Coral reefs protect coastlines from storm surge and support commercial and recreational fisheries and tourism. Unfortunately, certain chemicals in sunscreen are causing coral reefs to bleach; thus, becoming more susceptible to viral infections. The reefs eventually turn white and die. Coral reef bleaching is the leading cause of coral reef deaths worldwide. This conversation is an important one to discuss leading up to the celebration of World Oceans Day on June 8.

Chemical recreational sunscreen contains oxybenzone, a toxic synthetic molecule. Oxybenzone is prevalent in the majority of mainstream sunscreen brands. This ingredient results in extreme harm to marine organisms. The Ocean Foundation emphasized that, “A single drop of this compound in more than 4 million gallons of water is enough to endanger organisms.” Even if you do not physically go in the water, the chemical can be washed into the ocean through the sand.

In response to this issue, many countries and resorts are banning “reef-toxic” sunscreen. Hawaii and Key West recently passed a bill banning the sale and distribution of any sunscreen that contains 10 toxic ingredients, including oxybenzone. This bill goes into effect January 2021. Many dermatologists are concerned for public safety, highlighting that banning certain sunscreens will decrease overall use. Unprotected sun exposure it the most preventable risk factor for skin cancer. From the perspective of a customer, it is important to be actively informed on what constitutes a “reef-safe” sunscreen. Oxybenzone can pop-up in many moisturizers, primers, and foundations that contain SPF. Reef-friendly options include: any version of chemical sunscreen that does not contain oxybenzone.

With a commitment to protect the environment, Promega has pledged $3 million over the next three years to the Revive and Restore Catalyst Science fund. Organization founders and scientists are focused on an extremely long-term view of wildlife conservation. This fund invests in proof-of-concept research projects that offer innovative solutions for conservation challenges and threatened ecosystems. Marine biologist Steve Palumbi was awarded the first Fund grant to investigate the triggers that may cause corals to bleach. Palumbi reflects on his research in an interview with Stanford News stating, “The report reflects a sense of urgency. We need to start helping corals now, so that as the climate gets worse—and it will inevitably get worse—we’re a little bit in front of the problem. There’s this amazing sense that we all have to just jump in and try ideas and fail so that, eventually, someone comes up with the answers we need.”

Announcing the 2019 Promega iGEM Grant Winners

Posted on by Jordan Villanueva

It’s FINALLY time to announce the winners of the 2019 Promega iGEM Grant! We received over 150 applications this year, so picking the top 10 was very tough. As always, we’re impressed by the amazing work iGEM teams are doing in the lab and in their communities. The 10 winners listed below will receive $2,000 in free Promega products.

Good luck to all teams competing in iGEM this year, and congratulations to our winners! Don’t forget that Promega has free technical support for all teams competing in iGEM. Our scientists are excited to help out. You can also check out our iGEM Sponsor page, which has tools and resources to help make your project a success. Continue reading “Announcing the 2019 Promega iGEM Grant Winners”

Why You Don’t Need to Select a Wavelength for a Luciferase Assay

Posted on by Ben Schmidt
Promega kit depicted; test involves wavelength for a luciferase assay.

It’s a question I’m asked probably once a week. “What wavelength do I select on my luminometer when performing a luciferase assay?” The question is a good and not altogether unexpected one, especially for those new to bioluminescent assays. The answer is that in most cases, you don’t and in fact shouldn’t select a wavelength (the exception to this rule is if you’re measuring light emitted in two simultaneous luciferase reactions). To understand why requires a bit of an explanation of absorbance, fluorescence, and luminescence assays, and the differences among them.

Absorbance, fluorescence, and luminescence assays are all means to quantify something of interest, be that a genetic reporter, cell viability, cytotoxicity, apoptosis, or other markers. In principle, they are all similar. For example, a genetic reporter assay is an indicator of gene expression. The promoter of a gene of interest can be cloned upstream of a reporter such as β-galactosidase, GFP, or firefly luciferase. The amount of each of these reporters that is transcribed into mRNA and translated into protein by the cell is indicative of the endogenous expression of the gene of interest.

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The Simplex Things In Life: Utilizing Artificial Intelligence Models to Better Understand Autism

Posted on by Natalie Larsen

Autism Spectrum Disorder, or ASD, is nothing if not unique.

The way ASD manifests itself in people is unique; although it most often presents as some form of variable impairment in social interaction and communication, each individual has behaviors and habits that are as unique to them as snowflakes are to one another.

ASD has also proven itself to be a uniquely challenging disorder to study. In the past decade, de novo (new) mutations have been identified as key contributors to causality of ASD. However, the majority of these identified de novo mutations are located in protein-coding genes, which comprise only 1–2% of the entire human genome.

Up to this point, a majority of previous research has focused on identifying mutations located in the 20,000 identified genes in the protein-coding region, which would seem like a promising approach. Genes are the genetic blueprints for creating proteins, which control and perform crucial tasks in our bodies, such as fighting off infections, communicating between your organs, tissues, and cells as chemical messengers, and regulating your blood sugar levels. It seems like basic math: Genes + Mutations = Mutated Proteins. Mutated Proteins = Disrupted Protein Function.

However, it has been observed that all the known genes that are ASD-associated can explain only a minor fraction of new autism cases, and it is estimated that known de novo mutations in the protein-coding region contribute to not more than 30% of cases for individuals who have no family history of autism (better known as simplex ASD). This provides evidence to suggest mutations contributing to autism must additionally occur elsewhere in the genome. Continue reading “The Simplex Things In Life: Utilizing Artificial Intelligence Models to Better Understand Autism”

What Packaging Dog Food Can Teach You About Science Writing and Other Tales

Posted on by Kassidy Steyer

Cartoon drawing of two workers drinking coffee

Believe it or not, the most unglamorous jobs teach us all a thing or two about life. I asked Promega staff members to discuss the most impactful lessons they learned at their first jobs. Check it out.

Continue reading “What Packaging Dog Food Can Teach You About Science Writing and Other Tales”

When Proteins Get Together: Shedding (Blue) Light on Cellular LOV

Posted on by Ken Doyle

NanoBRETNo protein is an island. Within a cell, protein-protein interactions (PPIs) are involved in highly regulated and specific pathways that control gene expression and cell signaling. The disruption of PPIs can lead to a variety of disease states, including cancer.

Two general approaches are commonly used to study PPIs. Real-time assays measure PPI activity in live cells using fluorescent or luminescent tags. A second approach includes methods that measure a specific PPI “after the fact”; popular examples include a reporter system, such as the classic yeast two-hybrid system.

Continue reading “When Proteins Get Together: Shedding (Blue) Light on Cellular LOV”

What Makes a “Good” Buffer?

Posted on by Michele Arduengo
Use of buffers, pour one solution into another.
Use of buffer aims to make pH remain nearly constant in solution.

Buffers are often overlooked and taken for granted by laboratory scientists, until the day comes when a bizarre artifact is observed and its origin is traced to a bad buffer.

The simplest definition of a buffer is a solution that resists changes in hydrogen ion concentration as a result of internal and environmental factors. Buffers essentially maintain pH for a system. The effective buffering range of a buffer is a factor of its pKa, the dissociation constant of the weak acid in the buffering system. Many things, such as changes in temperature or concentration, can affect the pKa of a buffer.

In 1966, Norman Good and colleagues set out to define the best buffers for biochemical systems (1). By 1980, Good and his colleagues identified twenty buffers that set the standard for biological and biochemical research use (2,3).  Good set forth several criteria for the selection of these buffers:

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Evaluating CAR NK Immunotherapy in Patient-Derived Colorectal Organoids

Posted on by Johanna Lee

In recent years, great advances have been made in the field of immunotherapy to treat cancer. One of the most promising treatments involves engineering immune cells to express chimeric antigen receptors (CAR). These receptors are carefully designed to recognize antigens expressed on the surface of tumor cells. Once the target is recognized, the CAR-engineered immune cells can attack and kill the tumor cells. CAR T cells have been successfully used to treat certain blood cancers—three CAR T therapies for lymphoma and leukemia have gained US FDA approval. In these cases, T cells were taken from individual patients, grown and genetically-altered in the lab, then reintroduced into the same patient. Continue reading “Evaluating CAR NK Immunotherapy in Patient-Derived Colorectal Organoids”

Making Research More Sustainable, One Lab at a Time

Posted on by Darcia Schweitzer

Do you love your research job? What if you couldn’t do that work anymore? What if future researchers couldn’t have the opportunity to build from what you have accomplished and feel the same joy you do about their research?

Unfortunately, these may become more than hypotheticals for the next generation of scientists due to the impact humans are having on the earth. Scientific research has an outsized impact on some aspects of our unsustainable use of resources. Academic research buildings can use four times more energy than a typical office building and can be responsible for one-third of all waste generated on campus. So, can you make scientific research more sustainable?

Continue reading “Making Research More Sustainable, One Lab at a Time”