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技術(shù)文章

*脫羧酶2(GAD2)檢測試劑盒

閱讀:527發(fā)布時間:2015-5-7

*脫羧酶2(GAD2)檢測試劑盒

適用生物 Homo sapiens (Human,人)
*脫羧酶2(GAD2)檢測試劑盒檢測范圍 0.781-50ng/mL 靈敏度 0.33ng/mL
樣本類型 Tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
實驗時長 4.5h 實驗方法 雙抗夾心法 *脫羧酶2(GAD2)檢測試劑盒規(guī)格 96T

ELISA Kit for Glutamate Decarboxylase 2, Acid (GAD2)

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

Organism speciesHomo sapiens (Human)
Product No.hzEB258Hu
Sample typeTissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Format96-well strip plate
Assay length4.5 hours
Detection range0.781-50ng/mL The standard curve concentrations used for the ELISA’s were 50ng/mL, 25ng/mL, 12.5ng/mL, 6.25ng/mL, 3.125ng/mL, 1.563ng/mL, 0.781ng/mL
SensitivityThe minimum detectable dose of this kit is typically less than 0.33ng/mL.

*脫羧酶2(GAD2)檢測試劑盒Specificity

This assay has high sensitivity and excellent specificity for detection of Glutamate Decarboxylase 2, Acid (GAD2).
No significant cross-reactivity or interference between Glutamate Decarboxylase 2, Acid (GAD2) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutamate Decarboxylase 2, Acid (GAD2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glutamate Decarboxylase 2, Acid (GAD2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

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.

*脫羧酶2(GAD2)檢測試劑盒Reagents and materials provided

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×120μLAssay Diluent A1×12mL
Detection Reagent B1×120μLAssay Diluent B1×12mL
TMB Substrate1×9mLStop Solution1×6mL
Wash Buffer (30 × concentrate)1×20mLInstruction manual1

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.

*脫羧酶2(GAD2)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 Glutamate Decarboxylase 2, Acid (GAD2). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Glutamate Decarboxylase 2, Acid (GAD2). 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 Glutamate Decarboxylase 2, Acid (GAD2), 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 Glutamate Decarboxylase 2, Acid (GAD2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.


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