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Multiplex Assay Kit for Low Density Lipoprotein Receptor Related Protein 1 (LRP1) ,etc. by FLIA (Flow Luminescence Immunoassay)

CD91; LRP; A2MR; APOER; APR; TGFBR5; Alpha-2-Macroglobulin Receptor; Prolow-density lipoprotein receptor-related protein 1; Apolipoprotein E receptor

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

Specificity

This assay has high sensitivity and excellent specificity for detection of Low Density Lipoprotein Receptor Related Protein 1 (LRP1).
No significant cross-reactivity or interference between Low Density Lipoprotein Receptor Related Protein 1 (LRP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Low Density Lipoprotein Receptor Related Protein 1 (LRP1) and the recovery rates were calculated by comparing the measured value to the expected amount of Low Density Lipoprotein Receptor Related Protein 1 (LRP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-103 89
EDTA plasma(n=5) 91-105 99
heparin plasma(n=5) 85-101 90
sodium citrate plasma(n=5) 80-101 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Low Density Lipoprotein Receptor Related Protein 1 (LRP1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Low Density Lipoprotein Receptor Related Protein 1 (LRP1) 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 Low Density Lipoprotein Receptor Related Protein 1 (LRP1) 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) 93-101% 79-99% 82-98% 78-97%
EDTA plasma(n=5) 85-105% 98-105% 79-97% 97-105%
heparin plasma(n=5) 99-105% 80-94% 99-105% 99-105%
sodium citrate plasma(n=5) 97-105% 80-88% 85-101% 83-105%

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#:LRP1) 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 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
European Journal of Vascular and Endovascular Surgery Plasma Low-density Lipoprotein Receptor-related Protein 1 Concentration is not Associated with Human Abdominal Aortic Aneurysm Presence Science: Article
Int J Cardiol Circulating soluble low-density lipoprotein receptor-related protein 1 (sLRP1) concentration is associated with hypercholesterolemia: A new potential biomarker for atherosclerosis PubMed: 26285183
International Journal of Cardiology Circulating levels of soluble low-density lipoprotein receptor-related protein 1 (sLRP1) as novel biomarker of epicardial adipose tissue pubmed:27543712
Journal of Cellular and Molecular Medicine Extracellular vesicles do not contribute to higher circulating levels of soluble LRP1 in idiopathic dilated cardiomyopathy doi:10.1111
Scientific Reports Soluble LRP1 is an independent biomarker of epicardial fat volume in patients with type 1 diabetes mellitus Pubmed:29348672
Atherosclerosis Soluble low-density lipoprotein receptor-related protein 1 as a biomarker of coronary risk: Predictive capacity and association with clinical events
Biomedical Reports Gender difference and genetic variance in lipoprotein receptor‑related protein 1 is associated with mortality
biorxiv. CMTM6 drives cisplatin resistance in OSCC by regulating AKT mediated Wnt signaling
JCI Insight CMTM6 drives cisplatin resistance by regulating Wnt signaling through ENO-1/AKT/GSK3¦Â axis 33434185
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