Ⅳ型胶原（COL4）检测试剂盒说明书
Ⅳ型胶原(COL4)检测试剂盒适用生物 Mus musculus (Mouse，小鼠) 检测范围 0.781-50ng/mL 灵敏度 0.37ng/mL 样本类型 Serum, plasma and other biological fluids 实验时长 4.5h 实验方法 双抗夹心法 规格 96T Ⅳ型胶原(COL4)检测试剂盒ELISA Kit for Collagen Type IV (COL4) FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species Mus musculus (Mouse) Product No. SEA180Mu Sample type Serum, plasma and other biological fluids Format 96-well strip plate Assay length 4.5 hours Detection range 0.781-50ng/mL The standard curve concentrations used for the ELISA’s were 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.125ng/mL, 1.563ng/mL, 0.781ng/mL Sensitivity The minimum detectable dose of this kit is typically less than 0.37ng/mL.
Ⅳ型胶原(COL4)检测试剂盒Specificity This assay has high sensitivity and excellent specificity for detection of Collagen Type IV (COL4). No significant cross-reactivity or interference between Collagen Type IV (COL4) and analogues was observed. Recovery Matrices listed below were spiked with certain level of recombinant Collagen Type IV (COL4) and the recovery rates were calculated by comparing the measured value to the expected amount of Collagen Type IV (COL4) in samples. Ⅳ型胶原(COL4)检测试剂盒
Matrix Recovery range (%) Average(%) serum(n=5) 88-102 96 EDTA plasma(n=5) 86-102 93 heparin plasma(n=5) 80-89 81
Precision Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Collagen Type IV (COL4) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Collagen Type IV (COL4) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: CVⅣ型胶原(COL4)检测试剂盒
Sample 1:2 1:4 1:8 1:16 serum(n=5) 88-104% 85-99% 80-93% 79-91% EDTA plasma(n=5) 82-96% 80-99% 83-101% 97-105% heparin plasma(n=5) 78-98% 88-96% 86-95% 93-101%
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 TMB Substrate 1×9mL Stop Solution 1×6mL Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1
Ⅳ型胶原(COL4)检测试剂盒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. 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 Collagen Type IV (COL4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Collagen Type IV (COL4). 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 Collagen Type IV (COL4), 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 Collagen Type IV (COL4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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