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Promega Corporation

4 ARTICLES PUBLISHED IN JoVE

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Biology

Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
Danette L. Daniels 1, Jacqui Méndez 1, Hélène Benink 1, Andrew Niles 1, Nancy Murphy 1, Michael Ford 2, Richard Jones 2, Ravi Amunugama 2, David Allen 2, Marjeta Urh 1
1Promega Corporation, 2MS Bioworks LLC

HaloTag technology is a multifunctional technology which has shown significant success in isolation of both small and large protein complexes from mammalian cells.  Here we highlight the advantages of this technology compared to existing alternatives and demonstrate its utility to study numerous aspects of protein function inside eukaryotic cells.

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Bioengineering

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo
Brian Burkel 1,2, Brett A. Morris 1, Suzanne M. Ponik 1, Kristin M. Riching 1, Kevin W. Eliceiri 2,3,4, Patricia J. Keely 1,2,5
1Department of Cell and Regenerative Biology, University of Wisconsin-Madison, 2Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, 3Department of Biomedical Engineering, University of Wisconsin-Madison, 4Morgridge Institute for Research, University of Wisconsin-Madison, 5Paul P. Carbone Comprehensive Cancer center, University of Wisconsin-Madison

Collagen is a core component of the ECM, and provides essential cues for several cellular processes ranging from migration to differentiation and proliferation. Provided here is a protocol for embedding cells within 3D collagen hydrogels, and a more advanced technique for generating randomized or aligned collagen matrices using PDMS microchannels.

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Immunology and Infection

Antibody Labeling with Fluorescent Dyes Using Magnetic Protein A and Protein G Beads
Nidhi Nath 1, Becky Godat 1, Marjeta Urh 1
1Promega Corporation

The on-bead method for labeling antibodies with small molecules enables labeling of a small amount of antibodies directly from cell media. This method is compatible with amine and thiol chemistry, and can handle multiple samples in parallel, manually or using automated platforms.

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Cancer Research

High-Throughput Cellular Profiling of Targeted Protein Degradation Compounds Using HiBiT CRISPR Cell Lines
Kristin M. Riching 1, Sarah D. Mahan 1, Marjeta Urh 1, Danette L. Daniels 1
1Promega Corporation

This protocol describes the quantitative luminescent detection of protein degradation kinetics in living cells that have been engineered using CRISPR/Cas9 to express antibody free endogenous protein detection tag fused to a target protein. Detailed instructions for calculating and obtaining quantitative degradation parameters, rate, Dmax, DC50, and Dmax50 are included.

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