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腎上腺髓質(zhì)素(ADM)檢測試劑盒
適用生物 Canis familiaris; Canine (Dog,犬)
腎上腺髓質(zhì)素(ADM)檢測試劑盒檢測范圍 12.35-1000pg/mL 靈敏度 4.42pg/mL
樣本類型 Serum, plasma and other biological fluids.
實驗時長 2.5h 實驗方法 競爭抑制法 腎上腺髓質(zhì)素(ADM)檢測試劑盒規(guī)格 96T
ELISA Kit for Adrenomedullin (ADM)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species Canis familiaris; Canine (Dog)
腎上腺髓質(zhì)素(ADM)檢測試劑盒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 4.42pg/mL.
Specificity
This assay has high sensitivity and excellent specificity for detection of Adrenomedullin (ADM).
No significant cross-reactivity or interference between Adrenomedullin (ADM) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Adrenomedullin (ADM) and the recovery rates were calculated by comparing 腎上腺髓質(zhì)素(ADM)檢測試劑盒the measured value to the expected amount of Adrenomedullin (ADM) in samples.
Matrix Recovery range (%) Average(%)
serum(n=5) 82-97 87
EDTA plasma(n=5) 92-105 98
heparin plasma(n=5) 85-96 89
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adrenomedullin (ADM) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adrenomedullin (ADM) 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 Adrenomedullin (ADM) 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) 88-102% 84-105% 89-105% 80-101%
EDTA plasma(n=5) 82-102% 98-105% 95-103% 88-102%
heparin plasma(n=5) 93-105% 90-103% 79-91% 97-105%
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 腎上腺髓質(zhì)素(ADM)檢測試劑盒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 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腎上腺髓質(zhì)素(ADM)檢測試劑盒 competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Adrenomedullin (ADM) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Adrenomedullin (ADM) and unlabeled Adrenomedullin (ADM) (Standards or samples) with the pre-coated antibody specific to Adrenomedullin (ADM). 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 Adrenomedullin (ADM) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the腎上腺髓質(zhì)素(ADM)檢測試劑盒 concentration of Adrenomedullin (ADM) in the sample.
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