ELISA Kit for Cathepsin L (CTSL)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 86-96 89
EDTA plasma(n=5) 82-98 91
heparin plasma(n=5) 80-96 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathepsin L (CTSL) 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) 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) 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) 90-105% 94-102% 79-98% 83-91%
EDTA plasma(n=5) 85-103% 95-102% 99-105% 88-101%
heparin plasma(n=5) 81-103% 82-101% 88-95% 78-91%

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 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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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