激活蛋白C（APC）检测试剂盒

【资料简介】

激活蛋白C(APC)检测试剂盒

适用生物 Homo sapiens (Human，人)
激活蛋白C(APC)检测试剂盒检测范围 31.25-2000pg/mL 灵敏度 13.1pg/mL
样本类型 Plasma
实验时长 4.5h 实验方法 双抗夹心法激活蛋白C(APC)检测试剂盒规格 96T
ELISA Kit for Activated Protein C (APC)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!

Organism species Homo sapiens (Human)
激活蛋白C(APC)检测试剂盒Sample type Plasma
Format 96-well strip plate
Assay length 4.5 hours
Detection range 31.25-2000pg/mL The standard curve concentrations used for the ELISA’s were 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL
Sensitivity The minimum detectable dose of this kit is typically less than 13.1pg/mL.

Specificity
This assay has high sensitivity and激活蛋白C(APC)检测试剂盒 excellent specificity for detection of Activated Protein C (APC).
No significant cross-reactivity or interference between Activated Protein C (APC) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Activated Protein C (APC) and the recovery rates were calculated by 激活蛋白C(APC)检测试剂盒comparing the measured value to the expected amount of Activated Protein C (APC) in samples.

Matrix Recovery range (%) Average(%)
sodium citrate plasma(n=5) 92-99 96

Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Activated Protein C (APC) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Activated Protein C (APC) 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激活蛋白C(APC)检测试剂盒 by testing samples spiked with appropriate concentration of Activated Protein C (APC) 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
sodium citrate plasma(n=5) 97-105% 92-99% 93-101% 79-89%

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激活蛋白C(APC)检测试剂盒 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 37oC;
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37oC;
4. Aspirate and wash 3 times;
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
6. Aspirate and wash 5 times;
7. Add 90μL Substrate Solution.激活蛋白C(APC)检测试剂盒 Incubate 15-25 minutes at 37oC;
8. Add 50μL Stop Solution. Read at 450nm immediay.
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 Activated Protein C (APC). Standards or samples are then added 激活蛋白C(APC)检测试剂盒to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Activated Protein C (APC). 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 Activated Protein C (APC), 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激活蛋白C(APC)检测试剂盒 the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Activated Protein C (APC) in the samples is then determined by comparing the O.D. of the samples to the standard curve.