褪黑素(MT)检测试剂盒

褪黑素(MT)检测试剂盒

适用生物     General，通用    

褪黑素(MT)检测试剂盒   

检测范围     12.35-1000pg/mL     灵敏度     4.94pg/mL    

样本类型     Serum, plasma and other biological fluids.    

实验时长     2.5h     实验方法     竞争抑制法    褪黑素(MT)检测试剂盒

规格     96T    

ELISA Kit for Melatonin (MT)

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

Organism species     General    

褪黑素(MT)检测试剂盒

Sample type     Serum, plasma and other biological fluids.    

Format     96-well strip plate    

Assay length     2.5 hours    

Detection range     12.35-1000pg/mL The standard curve concentrations used for the ELISA’s were 1000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL    

Sensitivity     The minimum detectable dose of this kit is typically less than 4.94pg/mL.    

Specificity

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

Recovery

Matrices listed below were spiked 褪黑素(MT)检测试剂盒with certain level of recombinant Melatonin (MT) and the recovery rates were calculated by comparing the measured value to the expected amount of Melatonin (MT) in samples.

Matrix     Recovery range (%)     Average(%)    

serum(n=5)     86-101     96    

EDTA plasma(n=5)     85-101     93    

heparin plasma(n=5)     87-94     90    

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Melatonin (MT) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Melatonin (MT) 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 Melatonin (MT) and their serial褪黑素(MT)检测试剂盒 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)     83-93%     80-93%     93-101%     96-103%    

EDTA plasma(n=5)     79-91%     78-96%     79-94%     99-105%    

heparin plasma(n=5)     81-103%     82-99%     80-101%     87-99%    

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     Assay Diluent A     1×12mL    

Detection Reagent B     1×120μL     Assay Diluent B     1×12mL    

Reagent Diluent     1×300μL     Stop Solution     1×6mL    

TMB Substrate     1×9mL     Instruction manual     1    

Wash Buffer (30 × concentrate)     1×20mL    

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50μL standard or sample to each well.
    And then add 50μL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37oC;
3. Aspirate and wash 3 times;
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50μL Stop Solution. Read at 450 nm immediately.

Test principle

This assay employs the 褪黑素(MT)检测试剂盒competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Melatonin (MT) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Melatonin (MT) and unlabeled Melatonin (MT) (Standards or samples) with the pre-coated antibody specific to Melatonin (MT). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Melatonin (MT) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the 褪黑素(MT)检测试剂盒concentration of Melatonin (MT) in the sample.