ELISA Kit for Cathepsin S (CTSS)

CTS-S

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 91-99 95
EDTA plasma(n=5) 88-103 91
heparin plasma(n=5) 91-99 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cathepsin S (CTSS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cathepsin S (CTSS) 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 S (CTSS) 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) 92-101% 88-96% 91-98% 89-96%
EDTA plasma(n=5) 80-101% 83-101% 86-101% 78-97%
heparin plasma(n=5) 80-93% 89-96% 84-102% 78-93%

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
Chinses Medical Journal Relationship between plasma cathepsin S and cystatin C levels and coronary plaque morphology of mild to moderate lesions: an in vivo study using intravascular ultrasound CMJ: 2009124346863507950
Journal of Cystic Fibrosis Protease–antiprotease imbalances differ between Cystic Fibrosis patients&#039; upper and lower airway secretions Pubmed:25286826
Scientific Reports Circulating cathepsin-S levels correlate with GFR decline and sTNFR1 and sTNFR2 levels in mice and humans. pubmed:28240259
Neuroscience The P2X7 receptor, cathepsin S and fractalkine in the trigeminal subnucleus caudalis signal persistent hypernociception in temporomandibular rat joints Pubmed: 30248434
TOXICON Modulatory effect of botulinum toxin type A on the microglial P2X7/CatS/FKN activated-pathway in antigen-induced arthritis of the temporomandibular joint of … Pubmed: 32882256
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