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激素敏感性脂肪酶(LIPE)檢測試劑盒
適用生物 Homo sapiens (Human,人)
激素敏感性脂肪酶(LIPE)檢測試劑盒檢測范圍 0.156-10ng/mL 靈敏度 0.065ng/mL
樣本類型 Serum, plasma and other biological fluids
實驗時長 4.5h 實驗方法 雙抗夾心法 激素敏感性脂肪酶(LIPE)檢測試劑盒規(guī)格 96T
ELISA Kit for Lipase, Hormone Sensitive (LIPE)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species | Homo sapiens (Human) |
Product No. | xyEB276Hu |
Sample type | Serum, plasma and other biological fluids |
Format | 96-well strip plate |
Assay length | 4.5 hours |
Detection range | 0.156-10ng/mL The standard curve concentrations used for the ELISA’s were 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.313ng/mL, 0.156ng/mL |
Sensitivity | The minimum detectable dose of this kit is typically less than 0.065ng/mL. |
Specificity
This assay has high sensitivity and excellent specificity for detection of Lipase, Hormone Sensitive (LIPE).
No significant cross-reactivity or interference between Lipase, Hormone Sensitive (LIPE) and analogues was observed.
激素敏感性脂肪酶(LIPE)檢測試劑盒Recovery
Matrices listed below were spiked with certain level of recombinant Lipase, Hormone Sensitive (LIPE) and the recovery rates were calculated by comparing the measured value to the expected amount of Lipase, Hormone Sensitive (LIPE) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 89-104 | 98 |
EDTA plasma(n=5) | 92-101 | 97 |
heparin plasma(n=5) | 84-105 | 95 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Lipase, Hormone Sensitive (LIPE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Lipase, Hormone Sensitive (LIPE) 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 Lipase, Hormone Sensitive (LIPE) 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) | 81-103% | 89-97% | 80-93% | 79-90% |
EDTA plasma(n=5) | 82-99% | 96-105% | 82-89% | 88-96% |
heparin plasma(n=5) | 98-105% | 95-102% | 87-94% | 90-98% |
激素敏感性脂肪酶(LIPE)檢測試劑盒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 immediay.
激素敏感性脂肪酶(LIPE)檢測試劑盒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 Lipase, Hormone Sensitive (LIPE). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Lipase, Hormone Sensitive (LIPE). 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 Lipase, Hormone Sensitive (LIPE), 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 Lipase, Hormone Sensitive (LIPE) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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