α-内啡肽（αEP）检测试剂盒

α-内啡肽(αEP)检测试剂盒

适用生物 Homo sapiens (Human，人)
α-内啡肽(αEP)检测试剂盒检测范围 12.35-1000pg/mL 灵敏度 4.45pg/mL
样本类型 Serum, plasma, tissue homogenates, cell culture supernates and other biological fluid.
实验时长 2.5h 实验方法 竞争抑制法α-内啡肽(αEP)检测试剂盒规格 96T
ELISA Kit for Alpha-Endorphin (aEP)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!

Organism species Homo sapiens (Human)
α-内啡肽(αEP)检测试剂盒Sample type Serum, plasma, tissue homogenates, cell culture supernates and other biological fluid.
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 4.45pg/mL.

Specificity
This assay has high sensitivity and excellent specificity for detection of Alpha-Endorphin (aEP).
No significant cross-reactivity or interference between Alpha-Endorphin (aEP) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Alpha-Endorphin (aEP) and the recovery rates were calculated by comparing the measured value to the expected amount of Alpha-Endorphin (aEP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 90
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 78-94 88

Precision
Intra-assay Precision (Precision α-内啡肽(αEP)检测试剂盒within an assay): 3 samples with low, middle and high level Alpha-Endorphin (aEP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Alpha-Endorphin (aEP) 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 Alpha-Endorphin (aEP) and theirα-内啡肽(αEP)检测试剂盒 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) 89-101% 84-94% 97-104% 78-93%
EDTA plasma(n=5) 93-101% 78-99% 86-101% 84-103%
heparin plasma(n=5) 97-105% 89-105% 78-90% 91-101%

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

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 immediay.
    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 immediay.
Test principle
This assay employs the competitive α-内啡肽(αEP)检测试剂盒inhibition enzyme immunoassay technique. A monoclonal antibody specific to Alpha-Endorphin (aEP) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Alpha-Endorphin (aEP) and unlabeled Alpha-Endorphin (aEP) (Standards or samples) with the pre-coated antibody specific to Alpha-Endorphin (aEP). 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 Alpha-Endorphin (aEP) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to theα-内啡肽(αEP)检测试剂盒 concentration of Alpha-Endorphin (aEP) in the sample.