Multiplex Assay Kit for Lysophosphatidylcholine (LPC) ,etc. by CBA (Cytometric Bead Array)

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Lysophosphatidylcholine (LPC) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Lysophosphatidylcholine (LPC) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-91 88
EDTA plasma(n=5) 84-99 93
heparin plasma(n=5) 95-102 98

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lysophosphatidylcholine (LPC) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Lysophosphatidylcholine (LPC) ,etc. by CBA (Cytometric Bead Array) 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) ,etc. by CBA (Cytometric Bead Array) 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) 88-101% 78-92% 94-102% 96-103%
EDTA plasma(n=5) 90-97% 80-90% 82-98% 87-95%
heparin plasma(n=5) 84-93% 88-102% 98-105% 83-103%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:LPC) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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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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