| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Biological |
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Overview
For quantitative determination of phospholipase D activity and evaluation of drug effects on phospholipase D metabolism. The assay uses OD570nm, or FL530/585nm for signal readout. Compatible sample input includes Biological. Typical stated assay timing is 30 min.
Key elements and design rationale
- Readout format: OD570nm, or FL530/585nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Biological, which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 0.04 U/L for interpreting low-signal samples.
- Feature emphasis: Sensitive. Use 10 µL samples. Detection range: colorimetric assay 0.06 – 10 U/L, fluorimetric assay 0.04 – 1 U/L.
Additional feature notes highlight Simple and High-throughput: the assay involves the addition of a single working reagent and can be readily adapted to high-throughput assays for drug screening. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of phospholipase d 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
Phospholipase D (PLD) catalyses the hydrolysis of the phosphodiester bond of glycerophospholipids to generate phosphatidic acid and a free headgroup. Abnormalities in PLD expression have been associated with human cancers. BioAssay Systems method provides a simple and high-throughput assay for measuring PLD activity. In this assay, PLD hydrolyzes phosphatidylcholine to choline which is determined using choline oxidase and an H2O2-specific dye. The optical density of the pink-colored product at 570nm or fluorescence intensity (530/585 nm) is directly proportional to the PLD activity in the sample.
Detection method
Colorimetric (OD 570 nm) or Fluorescent (FL 530/585 nm).
Detection limit and analytical sensitivity
Reported detection limit: 0.04 U/L.
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 phospholipase d in biological by OD570 nm, or FL530/585 nm readout.
- Compare treatment or phenotype groups using matched biological handling.
- Monitor time-course or pre/post changes in biological 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.
I was curious how much PLD protein was used per well to generate the two data graphs shown in the product information for the kit.
The specific activity of an enzyme in terms of U/mg varies with the preparation and can also decrease over time. Our current phospholipase D (from Streptomyces sp.) has a specific activity of 6 U/mg. So if we want to reproduce the graph on the datasheet, we need to use 1.67 mg of our phospolipase D to obtain the 10/UL data point. The specific activity of your enzyme preparation may be much lower (or higher) and you should to titrate the preparation starting at the highest possible concentration.
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.
The role of phospholipase D1 in liver fibrosis induced by dimethylnitrosamine in vivo
Zhu X et al (2014). The role of phospholipase D1 in liver fibrosis induced by dimethylnitrosamine in vivo. Dig Dis Sci. 59(8):1779-88. Assay: Phospholipase D in rat tissue.
