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ELISA Kit for Diacylglycerol (DAG)

DG; Diglyceride

Specificity

This assay has high sensitivity and excellent specificity for detection of Diacylglycerol (DAG).
No significant cross-reactivity or interference between Diacylglycerol (DAG) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of Diacylglycerol (DAG) and the recovery rates were calculated by comparing the measured value to the expected amount of Diacylglycerol (DAG) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 81-89 85
EDTA plasma(n=5) 80-91 86
heparin plasma(n=5) 89-103 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Diacylglycerol (DAG) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Diacylglycerol (DAG) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Diacylglycerol (DAG) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 78-92% 88-95% 90-102% 85-101%
EDTA plasma(n=5) 82-105% 79-91% 79-103% 89-96%
heparin plasma(n=5) 93-101% 85-99% 78-101% 90-97%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Acta Pharmacologica Sinica Wogonin ameliorates lipotoxicity-induced apoptosis of cultured vascular smooth muscle cells via interfering with DAG-PKC pathway Nature: 2011120a
Psychoneuroendocrinology Maternal stress predicts altered biogenesis and the profile of mitochondrial proteins in the frontal cortex and hippocampus of adult offspring rats PubMed: 26143539
The Journal of Biological Chemistry Aryl Hydrocarbon Receptor Plays Protective Roles against High Fat Diet (HFD)-induced Hepatic Steatosis and the Subsequent Lipotoxicity via Direct Transcriptional ReCavia (Guinea pig )lation of Socs3 Gene Expression Pubmed:26865635
Journal of the American Heart Association Pigment Epithelium-Derived Factor (PEDF) Improves Ischemic Cardiac Functional Reserve Through Decreasing Hypoxic Cardiomyocyte Contractility Through PEDF Receptor (PEDF-R). pubmed:27413044
International Journal of Molecular Medicine PEDF protects cardiomyocytes by promoting FUNDC1‑mediated mitophagy via PEDF-R under hypoxic condition Pubmed:29512692
British Journal of Pharmacology Bouchardatine analogue alleviates NAFLD/NASH in high fat fed mice via blunting ATP synthase activity Pubmed: 31113010
Gut Akkermansia muciniphila ameliorates non-alcoholic fatty liver disease by L-aspartate via interaction with liver
Histochem Cell Biol The expression of diacylglycerol kinase isoforms α and ζ correlates with the progression of experimental autoimmune encephalomyelitis in rats 34312706
Catalog No. Related products for research use of Pan-species (General) Organism species Applications (RESEARCH USE ONLY!)
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