脫氧核糖核酸酶Ⅰ（DNASE1）檢測試劑盒

【資料簡介】

脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒

適用生物 Homo sapiens (Human，人)
脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒檢測范圍 78.13-5000pg/mL 靈敏度 34pg/mL
樣本類型 Serum, plasma, urine, saliva, seminal plasma and other biological fluids.
實驗時長 4.5h 實驗方法 雙抗夾心法 脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒規格 96T

ELISA Kit for Deoxyribonuclease I (DNASE1)

FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!

Organism species	Homo sapiens (Human)
Product No.	SEB127Hu
Sample type	Serum, plasma, urine, saliva, seminal plasma and other biological fluids.
Format	96-well strip plate
Assay length	4.5 hours
Detection range	78.13-5000pg/mL The standard curve concentrations used for the ELISA’s were 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.13pg/mL
Sensitivity	The minimum detectable dose of this kit is typically less than 34pg/mL.

Specificity

This assay has high sensitivity and excellent specificity for detection of Deoxyribonuclease I (DNASE1).
No significant cross-reactivity or interference between Deoxyribonuclease I (DNASE1) and analogues was observed.

脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒Recovery

Matrices listed below were spiked with certain level of recombinant Deoxyribonuclease I (DNASE1) and the recovery rates were calculated by comparing the measured value to the expected amount of Deoxyribonuclease I (DNASE1) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	87-96	90
EDTA plasma(n=5)	96-103	99
heparin plasma(n=5)	96-103	101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Deoxyribonuclease I (DNASE1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Deoxyribonuclease I (DNASE1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Deoxyribonuclease I (DNASE1) 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)	98-105%	79-105%	80-102%	98-105%
EDTA plasma(n=5)	96-103%	95-103%	83-103%	79-101%
heparin plasma(n=5)	93-104%	86-99%	89-96%	99-105%

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 2 hours at 37^oC;
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37^oC;
4. Aspirate and wash 3 times;
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37^oC;
6. Aspirate and wash 5 times;
7. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37^oC;
8. Add 50μL Stop Solution. Read at 450nm immediay.

脫氧核糖核酸酶Ⅰ(DNASE1)檢測試劑盒Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Deoxyribonuclease I (DNASE1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Deoxyribonuclease I (DNASE1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Deoxyribonuclease I (DNASE1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Deoxyribonuclease I (DNASE1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.