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ELISA Kit for Lysophosphatidylcholine (LPC)

lysoPC; Lysolecithin; 1-Palmitoyl-sn-Glycero-3-Phosphocholine

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 96-103 99
EDTA plasma(n=5) 90-102 99
heparin plasma(n=5) 78-102 88

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lysophosphatidylcholine (LPC) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Lysophosphatidylcholine (LPC) 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 Lysophosphatidylcholine (LPC) 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) 97-105% 78-98% 91-105% 82-98%
EDTA plasma(n=5) 97-104% 87-96% 94-102% 84-101%
heparin plasma(n=5) 89-103% 80-96% 82-95% 79-94%

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
Scientific Reports High-fat diet-induced acceleration of osteoarthritis is associated with a distinct and sustained plasma metabolite signature pubmed:28811491
Stem Cells Iron Homeostasis Determines Fate of Human Pluripotent Stem Cells Via Glycerophospholipids‐Epigenetic Circuit Pubmed: 30599084
Lipids in Health and Disease Associations between plasma lysophospholipids concentrations, chronic kidney disease and the type of renal replacement therapy Pubmed: 30947711
Nature Communications ER-residential Nogo-B accelerates NAFLD-associated HCC mediated by metabolic reprogramming of oxLDL lipophagy Pubmed: 31358770
NUTRIENTS Dietary Betaine Addition Promotes Hepatic Cholesterol Synthesis, Bile Acid Conversion, and Export in Rats Pubmed: 32414094
Cells iPLA2¦Â Contributes to ER Stress-Induced Apoptosis during Myocardial Ischemia/Reperfusion Injury
European Journal of Pharmacology Downregulation of activating transcription factor 4 attenuates lysophosphatidycholine-induced inflammation via the NF-κB pathway 34582847
Oxid Med Cell Longev Astragaloside IV Inhibits Bleomycin-Induced Ferroptosis in Human Umbilical Vein Endothelial Cells by Mediating LPC 34760046
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