Promega-Academy

Explore protein:protein interactions with the versatile platform technologies NanoBiT^® and NanoBRET™. Study the kinetics of reversible homo- and heterodimerization of proteins in live cells with these bioluminescent technologies. They accurately reflect the highly dynamic nature of protein:protein interactions.

You will learn:

• What is NanoBiT^®- and NanoBRET™-Technology
• How to study protein:protein interactions
• How to set up these assays in your lab

Combine CRISPR Knock-In technology with the HiBiT protein tagging system. This combination enables the construction of cellular reporter assays for a wide variety of research questions. The benefits of CRISPR & HiBiT are illustrated with examples of the research topics protein degradation and secretion, signal transduction and target cell killing.

You will learn:

• How to use CRISPR to engineer HiBiT-tagged proteins
• About live cell kinetic assays and endpoint assays
• How you can establish and use HiBiT-technology in your lab

Cell-based bioluminescent assays are standard methods for cell viability, cytotoxicity, and apoptosis measurement. The high sensitivity and broad linear range of these assays are fundamental to ensure their reproducibility. Despite choosing the right assay type, other factors need to be considered for a successful and reproducible implementation, ranging from plate selection to the choice of the correct detection mode.

You will learn:

• Tips and tricks for the optimal cell culture routine  
• How to find the best time point for analysis
• When to choose which detection mode  
• How to select the appropriate microtiter plate 
• Advantages of multiplexing and real-time assay formats  

Bioluminescent reporters are indispensable tools to study cellular signaling pathways and gene expression levels. In this seminar we will guide you through the key steps of a gene reporter experiment along with helpful tips and tricks. Applications of NanoLuc^® luciferase will prove the benefits of using NanoLuc^® luciferase for your reporter assays.

You will learn:

• How you can employ reporter assays in your research
• What the best reporter molecule is for your application
• The advantages of NanoLuc^® luciferase

Cell health assays provide valuable information about the viability of your cells. They are used for many applications including assessing the toxicity of drug candidates, analyzing cellular changes in cancer, or studying basic cell biology. This seminar provides an overview of the newest and most sensitive detection methods for cell viability, cytotoxicity, and apoptosis. 

The topics of this seminar are:

• How to set up the assays 
• Advantages and limitations influencing assay selection
• Kinetic assays for real-time assessment of cell health
• Assays optimized for 3D cell cultures
• How to get more data out of one sample 

This seminar shows you the possibilities to optimize your RT-qPCR workflow. Step by step, along the different work steps, tips and tricks will be shown and sources of errors will be revealed. From planning and performing the experiments, up to the correct analysis and interpretation of results, you will be an RT-qPCR expert at the end of this seminar!

You will learn:

• Tips & tricks for each step of RT-qPCR
• Correct sample handling and possibilities of RNA extraction
• Strategies of reverse transcription and correct enzyme selection
• Advantages and disadvantages of different qPCR systems
• Meaningful data analysis

Due to their high complexity, the development of antibody-based biologics presents an extraordinary challenge in drug development. Therefore, cell-based functional assays that reflect the mechanism of action (MOA) are required for potency evaluation, quality control, and manufacturing lot release. This seminar introduces state-of-the-art and innovative bioluminescent reporter bioassays for interrogating a range of biological functions:

• Fc-receptor mediated cellular response (ADCC and ADCP)
• Immune checkpoint modulation
• T-cell activation
• Cytokine and growth factor signaling

Cellular metabolism comprises a dynamic network of pathways that undergo reprogramming in response to external and internal signals. Metabolite levels provide valuable information about pathway activity. Plate-based coupled-enzyme reaction assays are rapid methods for measuring specific metabolites. This seminar provides an overview of simple, non-radioactive, bioluminescent methods for measuring metabolic changes in your cell culture.

In this seminar you will learn:

• Monitoring cellular key metabolites, e.g., glucose, lactate, glutamate 
• Oxidative stress detection
• Dinucleotide detection
• Monitoring lipid metabolism 

The new generation of immunoassays offers quantitative analyte detection and interaction analysis. Lumit™ Immunoassays are sensitive, have wide dynamic range and can be completed in as little as 30 minutes, making them a compelling alternative to labor-intensive ELISA and Western blot methodology.

You will learn:

• How Lumit™ Immunoassays simplify your workflows
• About various applications for Lumit™ Immunoassays in immunology, pharmacology, metabolism and SARS-CoV-2 research
• How you can build your own Lumit™ Immunoassays

• The basics of HaloTag^® technology
• How you can use HaloTag^® technology in your research
• About numerous applications including protein purification, cellular imaging or targeted protein degradation

The small and signal-intense NanoLuc^® luciferase serves as the starting point for the development of versatile protein reporter systems. Monitor the dynamics of protein:protein interactions with the NanoBiT^® protein tagging system. This technology led to the development of Lumit™ Immunoassays that enable fast quantification of proteins in ELISA-like immunoassays without washing steps.

You will learn:

• How to use NanoBiT^® and Lumit™ to study protein interactions
• How to quantify proteins at endogenous expression levels
• How to set up the platform technologies in your lab

A significant challenge is to characterize and optimize protein degraders (e.g., PROTACs, IMiDs, and molecular glues) for degradation efficacy. Currently, the availability of live-cell assays to interrogate the multiple steps required for targeted protein degradation is severely lacking. Here, we present a live-cell, luciferase-based technology platform that enables characterization of degrader compound mechanism of action.

You will learn:

• About bioluminescent reporter technology to study protein dynamics
• How to profit from either endpoint or real-time kinetic assay formats
• How you can quickly establish the technology in your lab