粘蛋白3(MUC3)检测试剂盒说明书

粘蛋白3(MUC3)检测试剂盒

适用生物     Homo sapiens (Human，人)    

粘蛋白3(MUC3)检测试剂盒    

检测范围     62.5-4000pg/mL     灵敏度     23.6pg/mL    

样本类型     Serum, plasma, tissue homogenates, cell lysates and other biological fluids.    

实验时长     4.5h     实验方法     双抗夹心法     粘蛋白3(MUC3)检测试剂盒

规格     96T    

粘蛋白3(MUC3)检测试剂盒       ELISA Kit for Mucin 3 (MUC3)

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

Organism species     Homo sapiens (Human)    

Product No.     SEB031Hu    

Sample type     Serum, plasma, tissue homogenates, cell lysates and other biological fluids.    

Format     96-well strip plate    

Assay length     4.5 hours    

Detection range     62.5-4000pg/mL The standard curve concentrations used for the ELISA’s were 4000pg/mL, 2000pg/mL, 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL    

Sensitivity     The minimum detectable dose of this kit is typically less than 23.6pg/mL.    

Specificity

This assay has high sensitivity and excellent specificity for detection of Mucin 3 (MUC3).
No significant cross-reactivity or interference between Mucin 3 (MUC3) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mucin 3 (MUC3) and the recovery rates were calculated by comparing the measured value to the expected amount of Mucin 3 (MUC3) in samples.

Matrix     Recovery range (%)     Average(%)    

serum(n=5)     97-105     101    

EDTA plasma(n=5)     96-104     99    

heparin plasma(n=5)     80-104     83    

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mucin 3 (MUC3) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mucin 3 (MUC3) 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 Mucin 3 (MUC3) 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)     97-104%     99-105%     85-101%     81-103%    

EDTA plasma(n=5)     82-89%     82-91%     86-95%     80-89%    

heparin plasma(n=5)     95-102%     92-99%     86-94%     94-103%    

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 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 immediately.

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 Mucin 3 (MUC3). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Mucin 3 (MUC3). 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 Mucin 3 (MUC3), 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 Mucin 3 (MUC3) in the samples is then determined by comparing the O.D. of the samples to the standard curve.