Using screening assays in biotechnology, the food industry and other fields relying on enzymes is time-consuming and often requires expert staff. GlycoSpot™ technology simplifies this process to a 3-step procedure that can be done within 30 minutes.
The technology can be utilized in several formats with minimal hands-on time, for example in a multiwell-plate format, which enables high-throughput enzyme activity screening of up to 96 samples at a time.
Apart from our GlycoSpot™ Multi-substrate enzyme screening assay kits we offer individual solutions. GlycoSpot™ Standard Assay kits have 20 Chromogenic Polymer Hydrogel (CPH) substrates available and we also offer Assay kits with 6 Insoluble Chromogenic Biomass (ICB) substrates. Furthermore, we are able to customize enzyme-screening solutions focused on your specific needs. Tell us what you are looking for and we will work out a solution tailored to you.
Chromogenic Polymer Hydrogel (CPH) substrates are substrates synthesized from pure polysaccharides and proteins. They are chemically dyed and crosslinked rendering them insoluble and chromogenic. Insoluble Chromogenic Biomass (ICB) substrates are substrates synthesized from raw plant material.
How do GlycoSpot™ assay kits work?
Upon digestion with enzymes that degrade the biopolymer(s) that the substrate was synthesized from, both CPH and ICB substrates release soluble dyed fragments into the incubation solution producing a colored supernatant.
Our substrates are arranged in a 96-well filter plate format, which enables high-throughput screening of up to 96 different reactions in a single plate.
After activation, enzyme solutions are added into the assay plate, incubated for the desired time at a temperature of your choice (stable up to 90 °C) and then filtered into a fresh 96-well receiving plate, where the activity can be assessed either by visual inspection or spectrophotometrically. The filtration can be achieved by either vacuum or centrifugation. A scheme of the process is shown below.
Our assay protocol consists of 3 simple steps:
How can GlycoSpot™ technology help my work?
GlycoSpot™ offers different high-throughput assay solutions to suit your specific needs. Multi CPH kits enable fast identification of a preferred substrate for an enzyme/enzyme mixture whose specificity is unknown. Standard CPH kits enable more detailed measurements of enzymes/enzyme mixtures with respect to identification of optimal conditions or batch variation assessments. ICB kits enable evaluation of how efficient and enzyme/enzyme mixture is when challenged with a substrate that very closely resembles complex biological material that an enzyme/enzyme mixture would encounter in nature. Additionally, Customized kits provide the customer with a possibility to design their own personalized assay kit with the layout and substrates that they require.
The Multi CPH kit
The Multi-Substrate CPH kit contains 16 different substrates that are perfect for pinpointing the preferred substrate of an enzyme/enzyme mixture whose specificity is unknown. This kit provides an excellent starting point in enzyme discovery and initial evaluation and quantification. The 16 substrates have been chosen to cover the most common and most abundant polysaccharides that are natural constituents of plant, bacterial and fungal cell walls together with polysaccharides with roles in energy storage.
The Standard CPH kit
The Standard CPH kit is comprised of the same CPH-substrate in all 96 multiplate wells and represents a valuable high-throughput solution for assessing enzyme concentration/activity/efficiency for enzymes/enzyme mixtures whose specificity is already known. Potential applications include batch-to-batch variation assessment in enzyme expression quality tracking, enzyme activity optimization with regard to different conditions such as pH, temperature and buffer components and many others.
The ICB substrate kit
ICB substrates are unique substrates produced from raw biological material (biomass). We have had great success in producing responsive ICB substrates from raw plant material, pre-treated biomass samples used in bioindustry and even biological material from other organisms such as animals, fungi and algae.
ICB substrates add an additional dimension to information gathered on a specific enzyme’s activity regarding its relative specificity/activity with respect to its target in pure form (CPH) or in a complex natural form (ICB).
The additional dimension arises from the fact that CPH substrates are synthesized from pure polysaccharides and ICB substrates from complex biological material. Activities recorded from a given enzyme towards a CPH or an ICB substrate therefore represent the activity of that enzyme towards a pure target or a target in its natural context, respectively. The natural context of ICB substrates implies that the polysaccharide/protein constituents are entangled in a complex network together with other compounds and may not be as accessible to the enzyme as when they are encountered alone as is the case with CPH substrates.
The Customized kit
With Customized kits we want to provide the customer with a possibility to design their own layouts of the assays with regard to the specific combinations of the substrates they want, including both CPH and ICB substrates. This also includes different substrate color options and customers can even, in the case of ICB substrates, send us a sample of their own biomass materials where we can attempt to produce a responsive substrate to suit their needs.
All GlycoSpot™ kits come with an information sheet that contains QC information with detailed measurements of absorbances of all substrates in the kit when tested with commercial enzymes providing our customers with a positive control reference point for the measurement they obtain in their experiments.
What is the difference between CPH and ICB substrates?CPH substrates are synthesized from pure polysaccharides and ICB substrates from complex biological material. Activities recorded from a given enzyme towards a CPH or an ICB substrate therefore represent the activity of that enzyme towards a pure target or a target in its natural context, respectively. The natural context of ICB substrates implies that the polysaccharide constituents are entangled in a complex network and may not be as accessible to the enzyme as when they are encountered alone in CPH substrates. This adds an additional dimension to information gathered on enzyme activity regarding its relative specificity/activity with respect to its target in pure form (CPH) or in a complex natural form (ICB).
A detailed protocol is delivered with all orders.
"A new generation of versatile chromogenic substrates for high-throughput analysis of biomass-degrading enzymes" by Kracun et al., 2015, published in Biotechnology for Biofuels. The methodology behind GlycoSpot's technology. The article can be accessed and dowloaded free of charge from the journal's own web page.