| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Compounds that affect glycolysis activity |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitative determination of L-Lactate and screening for glycolysis modulators. The assay uses OD565nm for signal readout. Compatible sample input includes Compounds that affect glycolysis activity. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD565nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Compounds that affect glycolysis activity, which is useful when aligning matrix type with calibration and control design.
- Workflow timing: The listed assay time of 30 min helps frame batch planning, replicate handling, and plate throughput.
- Feature emphasis: Fast and sensitive. Use of 5 µL sample. Linear detection range up to 10 mM L-lactate in 96-well plate assay.
Additional feature notes highlight Convenient. The procedure involves adding a single working reagent, and reading the absorbance after 30 minutes. Room temperature assay. No 37°C heater is needed; High-throughput. “Add-mix-read” type assay. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of glycolysis within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
GLYCOLYSIS is one of the major metabolic pathways cells undergo to produce energy and results in the production of pyruvate. One of the eventual fates of pyruvate from this process is lactate dehydrogenase converting it to L-lactate via lactic acid fermentation allowing L-lactate to serve as an indicator of glycolysis. BioAssay Systems Glycolysis assay kit is based on measuring the production of L-Lactate from glycolysis in cells. L-Lactate that is secreted into the cell media is quantified using a coupled reaction involving the lactate dehydrogenase catalyzed oxidation of L-lactate that generates pyruvate and NADH which reduces a formazan dye. The intensity of the reduced dye, measured at 565 nm, is directly proportional to the L-lactate concentration in the sample, which in turn is directly proportional to the glycolytic rate of the cell
Detection method
Colorimetric (OD 565 nm).
Procedures and timing
Stated procedure or timing information: 30 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- The description supports intervention-focused study designs in which researchers compare baseline and perturbed conditions.
Common research applications
- Quantify glycolysis in compounds that affect glycolysis activity by OD565 nm readout.
- Compare treatment or phenotype groups using matched compounds that affect glycolysis activity handling.
- Monitor time-course or pre/post changes in compounds that affect glycolysis activity across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
How do I store the kit?
This kit is shipped on ice. Upon receiving, please keep the kit in the freezer (-20°C).
What samples have you tested?
The kit has been tested on PANC-1, HL-60, F0-B16, and NIH-3T3 cell lines. It has been tested with DMEM and RPMI media. It has also been tested with 2-deoxy-D-glucose inhibition of glycolysis.
How do I prepare samples for assays?
For adherent cells, plate cells at desired cell density in media of choice. After allowing cells enough time to adhere to plate, remove media and replace with low percentage FBS media. Apply any treatments being tested at this time. Allow cells to propagate to desired confluency. Remove media for assay.
For suspension cells, seed cells in low percentage FBS media and apply any treatments being tested. Allow cells to grow to desired cell density. Remove media for assay.
Do suspension cells need to be spun down prior to assaying?
The kit has been tested on and works with suspension cells. Although it is not necessary to spin them down, we have found spinning them down prior to assay leads to more consistent results.
Does the assay work with 10% FBS media?
The assay does work with 10% FBS media; however, it loses a significant amount of sensitivity due to high background. If one chooses to run the assay with 10% FBS media, pilot experiments may be needed to determine optimal conditions. We strongly recommend simply using a low percentage FBS (FBS <1%) or serum free media.
My reader is only accurate to OD 1.0, can I use this assay on it?
The assay standards reach ODs greater than 1.0. If your reader is not accurate to OD 2.0, we suggest making a modified set of standards at concentrations 0, 1.5, 3, and 5mM. Samples may need to be diluted in media so they fall within this range and be corrected for dilution factor in calculations.
Do I need to use a standard or standard curve with each assay run?
Yes, it is highly recommended.
Can I store unused reagents for future use?
Yes, unused reagents can be stored according to the assay protocol. Excessive freeze/thaw cycles of reagents should be avoided.
For laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
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Umezu, R., et al. (2020). Macrophage (Drp1) dynamin-related protein 1 accelerates intimal thickening after vascular injury. Arteriosclerosis, Thrombosis, and Vascular Biology, 40(7), e214-e226. Assay: Glycolysis in mouse cell media.
Glucagon-like peptide-1 receptor agonism improves nephrotoxic serum nephritis by inhibiting t-cell proliferation
Moschovaki Filippidou, F., et al (2020). Glucagon-like peptide-1 receptor agonism improves nephrotoxic serum nephritis by inhibiting t-cell proliferation. The American Journal of Pathology, 190(2), 400-411 Assay: Glycolysis in mouse T cells.
Lysophosphatidic acid receptor activation affects the C13NJ microglia cell line proteome leading to alterations in glycolysis, motility, and cytoskeletal architecture
Bernhart, E., et al (2010). Lysophosphatidic acid receptor activation affects the C13NJ microglia cell line proteome leading to alterations in glycolysis, motility, and cytoskeletal architecture. Proteomics, 10(1), 141-158. Assay: Glycolysis in cell media.
Intermittent calorie restriction enhances epithelial-mesenchymal transition through the alteration of energy metabolism in a mouse tumor model
Kusuoka, Osamu, et al (2018). Intermittent calorie restriction enhances epithelial-mesenchymal transition through the alteration of energy metabolism in a mouse tumor model. International journal of oncology 52.2: 413-423. Assay: Glycolysis in mice cells.
Glycolysis-optimized conditions enhance maintenance of regenerative integrity in mouse spermatogonial stem cells during long-term culture
Helsel, Aileen R., Melissa J. Oatley, and Jon M. Oatley(2017). Glycolysis-optimized conditions enhance maintenance of regenerative integrity in mouse spermatogonial stem cells during long-term culture. Stem cell reports 8.5: 1430-1441. Assay: Glycolysis in mice cell culture.
