ELISA Kit for Cathepsin K (CTSK)

CTS-K; CTS02; CTSO; CTSO1; CTSO2; PKND; PYCD; Pycnodysostosis; Cathepsin O; Cathepsin X

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-102 87
EDTA plasma(n=5) 89-103 94
heparin plasma(n=5) 78-94 84

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathepsin K (CTSK) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cathepsin K (CTSK) 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 K (CTSK) 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) 82-90% 79-96% 99-105% 88-102%
EDTA plasma(n=5) 95-102% 98-105% 93-101% 98-105%
heparin plasma(n=5) 80-101% 91-104% 91-103% 96-105%

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
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
Acta Histochemica MRMT-1 rat breast carcinoma cells and models of bone metastases: improvement of an in vitro system to mimic the in vivo condition. PubMed: S0065128112000773
Atherosclerosis.? Biochemical and clinical correlation of intraplaque neovascularization using contrast-enhanced ultrasound of the carotid artery Pubmed:24534452
Genet. Mol. Res Drynaria total flavonoids decrease cathepsin K expression in ovariectomized rats Pubmed:25036175
Age (Dordr) In vitro method for the screening and monitoring of estrogen-deficiency osteoporosis by targeting peripheral circulating monocytes PubMed: 26250906
Journal of Infectious Diseases Cathepsin K Contributes to Cavitation and Collagen Turnover in Pulmonary Tuberculosis PubMed: 26416658
PLoS One. Effect of Vitamin D on Peripheral Blood Mononuclear Cells from Patients with Psoriasis Vulgaris and Psoriatic Arthritis pmc:PMC4822855
Cell Death And Differentiation TSC1 regulates osteoclast podosome organization and bone resorption through mTORC1 and Rac1/Cdc42 Pubmed:29358671
Indian Journal of Dental Research Comparative evaluation of cathepsin K levels in gingival crevicular fluid among smoking and nonsmoking patients with chronic periodontitis Pubmed: 30409937
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neuroscience letters Compound K induces neurogenesis of neural stem cells in thrombin induced nerve injury through LXRα signaling in mice Pubmed: 32371156
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