CLIA Kit for Sulfatase 2 (SULF2)

HSULF-2; Extracellular sulfatase Sulf-2

Specificity

This assay has high sensitivity and excellent specificity for detection of Sulfatase 2 (SULF2).
No significant cross-reactivity or interference between Sulfatase 2 (SULF2) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Sulfatase 2 (SULF2) and the recovery rates were calculated by comparing the measured value to the expected amount of Sulfatase 2 (SULF2) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-104 87
EDTA plasma(n=5) 91-104 96
heparin plasma(n=5) 78-89 82

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Sulfatase 2 (SULF2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Sulfatase 2 (SULF2) 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 Sulfatase 2 (SULF2) 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% 84-102% 83-101% 98-105%
EDTA plasma(n=5) 98-105% 96-104% 84-91% 97-105%
heparin plasma(n=5) 83-99% 83-97% 89-97% 83-101%

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
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
Substrate A 1×10mL Substrate B 1×2mL
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 100µL Substrate Solution. Incubate 10 minutes at 37°C;
8. Read RLU value immediately.

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Magazine Citations
Journal of Applied Pharmaceutical Science Cytotoxic effects of antiglypican-3 against HepG4 cell line Ebscohost: Source
European Journal of Pharmacology Evaluation of antiglypican-3 therapy as a promising target for amelioration of hepatic tissue damage in hepatocellular carcinoma Pubmed:25449037
Biomedicine & Pharmacotherapy Cytotoxic and partial hepatoprotective activity of sodium ascorbate against hepatocellular carcinoma through inhibition of sulfatase-2 and Pubmed:29669302
Experimental Dermatology Antitumor activity of sulforaphane in mice model of skin cancer via blocking sulfatase‐2 Pubmed: 30315662
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