Substrate Screening
Substrate Screening - Related Videos
11:13
Identification of Kinase-substrate Pairs Using High Throughput Screening
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Cited by 1 •
2015
Protein phosphorylation is a central feature of how cells interpret and respond to information in their extracellular milieu. Here, we present a high throughput screening protocol using kinases purified from mammalian cells to rapidly identify kinases that phosphorylate a substrate(s) of interest.
06:51
High-throughput Screening of Carbohydrate-degrading Enzymes Using Novel Insoluble Chromogenic Substrate Assay Kits
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Cited by 5 •
2016
A high-throughput assay for enzyme screening is described. This multiplexed ready-to-use assay kit comprises of pre-chosen Chromogenic Polymer Hydrogel (CPH) substrates and complex Insoluble Chromogenic Biomass (ICB) substrates. Target enzymes are polysaccharide degrading endo-enzymes and proteases.
03:49
Chromogenic Polymer Hydrogel-Based Enzyme Screening Assay: A High Throughput Method to Screen the Carbohydrate-Active Enzymes Using Synthetic Substrates
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2025
In this video, we demonstrate a high throughput assay for carbohydrate-active enzyme screening using insoluble chromogenic polymer hydrogel substrates. The enzyme degrades these substrates into a soluble colored product that can be quantified by measuring the color intensity of the product.
08:16
In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening
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2022
Here we present a protocol to express and purify recombinant Drosophila caspases Dronc and Drice, and their use in in vitro cleavage assays.
08:10
Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
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Cited by 7 •
2016
This work presents a method of high-throughput screening using a universal genetic enzyme screening system that can be theoretically applied to over 200 enzymes. Here, the single screening system identifies three different enzymes (lipase, cellulase, and alkaline phosphatase) by simply changing the substrate used (p-nitrophenyl acetate, p-nitrophenyl-β-D-cellobioside, and phenyl phosphate).
FAQs
Cells respond to differences in substrate composition, stiffness, roughness, wettability, and biochemical cues. These properties affect how cells interact with the material, which can change adhesion, spreading, viability, proliferation, or differentiation. By controlling these variables across candidate surfaces, researchers can connect particular material characteristics with specific biological responses and identify conditions that support the intended cell behavior.
These physical properties provide distinct ways to examine cell-substrate interactions. Stiffness, roughness, and wettability can be varied alongside composition or biochemical cues, allowing researchers to determine which material features correspond to stronger adhesion, broader spreading, improved viability, increased proliferation, or altered differentiation. Comparing them helps separate desirable biological effects from responses associated with another substrate characteristic.
No single measurement fully describes how cells perform on a material. Adhesion and spreading indicate how cells interact with the surface, while viability and proliferation show whether the environment supports cell maintenance and expansion. Differentiation provides information about changes in cell behavior. Evaluating these outcomes together helps researchers select substrates according to the biological performance required for a specific bioengineering application.
Researchers first select candidate surfaces or materials and establish controlled differences in properties such as composition, stiffness, roughness, wettability, or biochemical cues. They then culture cells on the candidates and measure relevant responses, including adhesion, spreading, viability, proliferation, or differentiation. Finally, they compare the results to identify materials associated with the desired biological behavior.
The measurement set can include cell adhesion, spreading, viability, proliferation, and differentiation. Together, these readouts describe whether cells attach to a surface, adopt a suitable spread state, remain viable, increase in number, or acquire a differentiated state. Selecting the most relevant outcomes allows the screening results to match the performance requirements of the planned material or culture platform.
Results from substrate screening guide the design of biomaterial coatings, scaffolds, and culture platforms with more predictable biological responses. The approach supports tissue engineering and regenerative medicine by helping identify materials compatible with desired cell behaviors. It also informs drug testing and other systems in which controlled cell-substrate interactions are important for interpreting or improving experimental performance.