BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
Store at 2-8°C,Protected from light,Do not freeze Ships Wet ice Check lot-specific COA for exact specifications.
SDS, COA, datasheet, and spec sheet available for download. Lot-specific COA accessible via lot number lookup.
Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Triglycerides (TG) are fat molecules formed by the condensation of three long-chain fatty acids with glycerol. They serve as crucial energy reserve substances and respiratory substrates in organisms.
Assay Principle:
Triglycerides are first hydrolyzed by lipoprotein lipase into glycerol and free fatty acids. Glycerol is then phosphorylated by glycerol kinase (GK) to produce glycerol‑1‑phosphate (G‑1‑P). Glycerol‑1‑phosphate is subsequently oxidized by glycerol‑3‑phosphate oxidase (GPO), generating hydrogen peroxide (H₂O₂). The hydrogen peroxide reacts with substrates such as 4‑aminoantipyrine to form a red quinone compound. This product has a characteristic absorption peak at 510 nm. By measuring the absorbance at this wavelength, the triglyceride content in the sample can be quantitatively determined.
Reagents, consumables and Equipments not provided
Procedure
It is recommended to first perform a pilot experiment using 1-3 samples with significant differences (e.g., different types or groups) to familiarize yourself with the procedure. Determine or adjust sample concentrations based on pilot results to avoid unnecessary waste of samples or reagents.
1. Sample Extraction
1.1 Tissue Samples
Weigh approximately 0.1 g of tissue and add it to a mortar with 1 mL of Extraction Solution. Homogenize on ice. Centrifuge at 12,000 rpm for 10 minutes at 4°C or room temperature. Collect the supernatant for assay.
Note: For high-fat tissue samples or partially high-fat samples, extraction with absolute ethanol is required.
1.2 Bacterial/Cell Samples
Collect bacteria or cells into a centrifuge tube, centrifuge, and discard the supernatant. Take approximately 5 million bacteria/cells and add 1 mL of Extraction Solution. Sonicate to disrupt the bacteria/cells (ice bath, 200 W power, 3 seconds sonication with 10-second intervals, repeated 30 times). Centrifuge at 12,000 rpm for 10 minutes at 4°C or room temperature. Collect the supernatant and keep it on ice for assay.
Note: To increase sample size, extraction can be performed at a ratio of 500~1000 (bacteria/cell count, ×10⁴) : 1 (Extraction Solution, mL).
1.3 Liquid Samples
Clear liquid samples can be assayed directly. If turbid, centrifuge and use the supernatant for detection.
2. Assay Procedure
2.1 Preheat the microplate reader for 30 minutes and set the wavelength to 510 nm.
2.2 Thaw all reagents to room temperature (25°C).
2.3 Add the following to a 96-well plate in order:
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Notes:
If A<sub>Assay</sub> > 0.3, dilute the sample (using Extraction Solution) or reduce the sample volume V1 (e.g., to 2 μL, with a corresponding increase in Reagent 1). The dilution factor D or the adjusted V1 must be used in the calculation formula.
For high-TG samples like serum or egg yolk (which requires ethanol extraction), sample volume V1 can be reduced to 2.5 μL and made up to 20 μL total with 17.5 μL distilled water, then proceed according to the table above. The adjusted V1 must be used in the calculation.
If (A<sub>Assay</sub> - A<sub>Blank</sub>) < 0.01, increase the sample volume V1 (e.g., from 5 μL to 10 μL, with a corresponding decrease in Reagent 1). The adjusted V1 must be used in the calculation formula.
3. Calculation of Results
3.1 Calculation Based on Sample Weight
TG (μg/g weight) = (CStd × V2) × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ (W × V1 ÷ V) × Mr × D
= CStd × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ W × Mr × D
3.2 Calculation Based on Cell Number
TG (μg/10⁴ cells) = (CStd × V2) × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ (500 × V1 ÷ V) × Mr × D
= CStd × (AAssay - ABlank) ÷ (AStd - ABlank) × 1.278 × D
3.3 Calculation of TG Content in Liquid
TG (μg/mL) = (CStd × V2) × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ V1 × Mr × D
= CStd × (AAssay - ABlank) ÷ (AStd - ABlank) × Mr × D
TG (mmol/L) = (CStd × V2) × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ V1 × D
= CStd × (AAssay - ABlank) ÷ (AStd - ABlank) × D
3.4 Calculation Based on Protein Concentration
TG (μg/mg prot) = (CStd × V2) × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ (Cpr × V1 ÷ V) × D
= CStd × (AAssay - ABlank) ÷ (AStd - ABlank) ÷ Cpr × Mr × D
Parameter Definitions:
CStd: Standard concentration 2.45mmol/L.
Mr: Molecular weight of triglyceride, 639.
V: Volume of Extraction Solution, 1 mL.
V1: Sample volume added, 0.005 mL.
V2: Standard volume added, 0.005 mL.
D: Dilution factor, 1 if not diluted.
W: Sample weight taken, g.
500: Cell number, 500 × 10⁴ cells.
AAssay: Absorbance of the assay well.
AStd: Absorbance of the standard well.
ABlank: Absorbance of the blank well.
Notes
Reagent 2 and the Standard are light-sensitive. Store and use protected from light at all times, avoiding direct sunlight.
For samples and reagents requiring ice bath operation, avoid repeated freeze-thaw cycles. Return unused reagents to 4°C storage promptly.
Aladdin's BCA Protein Quantification Kit (B665595) or Ready-to-Use BCA Protein Quantification Kit (R1491648) is recommended for protein concentration measurement.
| T1505523 | Component | 100T | Storage |
| T1505523A | Extraction Solution | 100 mL | 2-8℃. |
| T1505523B | Reagent I | 15 mL | 2-8℃. |
| T1505523C | Reagent II | 10 mL | 2-8℃. Store in the dark. |
| T1505523D | Standard (2.45mmol/L) | 1 EA | 2-8℃. Store in the dark |
Comprehensive hazard, handling, storage, and regulatory compliance document.
Download SDS →Lot-specific quality data. Enter your lot number to retrieve the exact COA.
Look up COA →Full quality attributes and acceptance criteria for this grade.
View spec sheet →Find and download the COA for your product by matching the lot number on the packaging.
| Lot Number | Certificate Type | Date | Item |
|---|---|---|---|
 ZJ26F0535652 | Certificate of Analysis | May 25, 2026 | T1505523 |
| ZJ26F0332662 | Certificate of Analysis | Mar 09, 2026 | T1505523 |
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