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調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)檢測試劑盒

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調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)檢測試劑盒

適用生物 Mus musculus (Mouse,小鼠)
調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)檢測試劑盒檢測范圍 78.13-5000pg/mL 靈敏度 34pg/mL
樣本類型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
實(shí)驗(yàn)時(shí)長 4.5h 實(shí)驗(yàn)方法 雙抗夾心法 調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)檢測試劑盒規(guī)格 96T

ELISA Kit for Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES)

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

Organism speciesMus musculus (Mouse)
Product No.hzEA116Mu
Sample typeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Format96-well strip plate
Assay length4.5 hours
Detection range78.13-5000pg/mL The standard curve concentrations used for the ELISA’s were 5000pg/mL, 2500pg/mL, 1250pg/mL, 625pg/mL, 312.5pg/mL, 156.25pg/mL, 78.13pg/mL
SensitivityThe minimum detectable dose of this kit is typically less than 34pg/mL.

Specificity

This assay has high sensitivity and excellent specificity for detection of Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES).
No significant cross-reactivity or interference between Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) and the recovery rates were calculated by comparing the measured value to the expected amount of Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) in samples.

MatrixRecovery range (%)Average(%)
serum(n=5)98-105101
EDTA plasma(n=5)85-9488
heparin plasma(n=5)81-9888

調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)檢測試劑盒Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) 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 Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample1:21:41:81:16
serum(n=5)81-101%88-96%80-102%81-97%
EDTA plasma(n=5)82-95%83-99%87-94%85-93%
heparin plasma(n=5)88-103%83-93%92-99%94-105%

調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)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

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.

調(diào)節(jié)激活正常T-細(xì)胞表達(dá)分泌因子(RANTES)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 Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES). 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 Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES), 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 Regulated On Activation In Normal T-Cell Expressed And Secreted (RANTES) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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