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Multiplex Assay Kit for Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay)

LTP; Lipid Transfer Protein; Plasma Lipid Transfer Protein; Cholesterylester Transfer Protein

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-97 93
EDTA plasma(n=5) 79-94 84
heparin plasma(n=5) 82-101 91

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Cholesteryl Ester Transfer Protein (CETP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 90-97% 78-90% 79-93% 80-91%
EDTA plasma(n=5) 90-101% 98-105% 96-103% 82-99%
heparin plasma(n=5) 78-105% 78-102% 83-95% 84-92%

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#:CETP) 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. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Neurotoxicology. 27-Hydroxycholesterol regulates cholesterol synthesis and transport in C6 glioma cells. pubmed:28167099
Nutrition A baru almond-enriched diet reduces abdominal adiposity and improves HDL concentrations: A randomized, placebo-controlled trial Pubmed:30086484
Scientific Reports CETP genetic variant rs1800777 (allele A) is associated with abnormally low HDL-C levels and increased risk of AKI during sepsis Pubmed: 30425299
Nutrition A baru almond└ enriched diet reduces abdominal adiposity and improves high-density lipoprotein concentrations: a randomized, placebo-controlled trial Pubmed: 30086484
Laboratory medicine Association of Serum Cholesterol Ester Transfer Protein Levels with Taq IB Polymorphism in Acute Coronary Syndrome Pubmed: 31504738
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