Multiplex Assay Kit for Cathepsin L (CTSL) ,etc. by FLIA (Flow Luminescence Immunoassay)

CTSL1; CATL; CTS-L; MEP; Cathepsin L1; Major excreted protein

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 84
EDTA plasma(n=5) 80-92 79
heparin plasma(n=5) 84-102 90

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathepsin L (CTSL) ,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 Cathepsin L (CTSL) ,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 Cathepsin L (CTSL) ,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) 96-105% 84-104% 93-104% 79-99%
EDTA plasma(n=5) 96-103% 85-93% 81-104% 93-101%
heparin plasma(n=5) 91-99% 83-105% 80-93% 94-101%

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#:CTSL) 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
Hypertension Identification of Cathepsin L as a Potential Sex-Specific Biomarker for Renal Damage. Ahajournals: source
The Journal of International Medical Research Plasma Cathepsin L and Its Related Pro/Antiangiogenic Factors Play Useful Roles in Predicting Rich Coronary Collaterals in Patients with Coronary Heart Disease Ingenta: art00021
Atherosclerosis.? Biochemical and clinical correlation of intraplaque neovascularization using contrast-enhanced ultrasound of the carotid artery Pubmed:24534452
International Journal of Molecular Sciences A New Analytical Method for Determination of Cathepsin L Based on the Surface Plasmon Resonance Imaging Biosensor Pubmed: 31052424
EMBO Molecular Medicine Immune‐mediated ECM depletion improves tumour perfusion and payload delivery Pubmed: 31709774
Cardiovasc Res Inhibition of myocardial cathepsin-L release during reperfusion following myocardial infarction improves cardiac function and reduces infarct size 34132807
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