BioCELL glycoTEST
Our BioCELL glycoTEST provides a precise measure of glycolysis in living intact cells, revealing transient responses and rapid metabolic switches not discernible in end-point lactate assays. Run on our Agilent Seahorse XF Analyzer, we quantify proton efflux specific to glycolysis.
How our test works.
Real-time measurements of glycolysis
A real-time assay to detect glycolytic modulation in minutes from living intact cells using XF analysis. Our clients use this assay to advance their understanding of immune cell activation, cancer cell metabolism and cell proliferation. The assay reports on key parameters, including basal glycolysis and compensatory glycolysis. Compensatory glycolysis is a measure of how flexible cells are in maintaining energy supply when mitochondrial respiration is inhibited. Compensatory glycolysis is a good indicator of how well a cell manages energy demand under stress.
An assay design backed by scientific evidence
Our assay measures baseline oxygen consumption rate (OCR) and proton efflux rate (PER) using XF analysis, followed by sequential injection of rotenone/antimycin A and 2-DG. Our data analysis removes the contribution of mitochondrial CO2 to PER using measurements of OCR, allowing accurate measurements of glycolytic-linked PER. It is well validated that glycolytic rate, as calculated from this assay, positively correlates with extracellular lactate efflux rate.
The gold-standard for studying glycolysis
Glycolysis and mitochondrial-derived CO2 are the two main contributors to extracellular acidification and proton efflux rate (PER) in cultured cells. The contribution of mitochondrial-derived CO2 to PER varies among different cell types, ranging from 3% to below 96%. Thus, it is critical to correct for the contribution of mitochondrial-derived CO2 to extracellular acidification to obtain accurate measurements of glycolysis. We apply this correction in our data analysis for both our BioCELL glycoTEST and BioCELL proFILE assays making them the best-in-class for establishing glycolytic function in living cells.
Key Features.
-
Real-time kinetic measurements
-
Live cell samples
-
Integrated injection ports
-
Label free sensors
-
Multi-parameter analysis
FAQs
-
This assay works for any cell line or primary cell type that uses aerobic glycolysis for maintaining energy demands. We can offer access to a range of different cell types including HEK293, HepG2, INS-1E, MCF-7, C2C12, PBMCs, T-lymphocytes, monocytes and macrophages.
-
Although measurements of oxygen consumption rate are assessed with this test, this is only to correct for mitochondrial contributions to CO2 acidification under basal conditions. This test is only designed to profile glycolytic function. If interested in mitochondrial function then see our BioCELL mitoTEST or our BioCELL proFILE assay, which have specifically been designed to profile mitochondrial function.
-
In some cases we would recommend optimising the assay, for example if a specific cell type or cell model is being used. We will also support any optimisation using our BioCELL OPTIMISE assay service.
-
Yes. We run a post-hoc fluorescent based nuclear stain cell density test. This is key to normalise any pre-treatment that might skew the results. Whereby this test is not suitable for the cell model being used we can also offer total protein or dsDNA for normalisation.
Interested in using our services?
Fill out the form below to arrange a free consultation with one of our experts.