适用生物 Rattus norvegicus (Rat,大鼠)
粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒
检测范围 7.81-500pg/mL 灵敏度 3.2pg/mL
样本类型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
实验时长 4.5h 实验方法 双抗夹心法
规格 96T
粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒
ELISA Kit for Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species |
Rattus norvegicus (Rat) |
Product No. |
SEA045Ra |
Sample type |
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
Format |
96T |
Assay length |
4.5 hours |
Detection range |
7.81-500pg/mL The standard curve concentrations used for the ELISA’s were 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.25pg/mL, 15.63pg/mL, 7.81pg/mL |
Sensitivity |
The minimum detectable dose of this kit is typically less than 3.2pg/mL. |
Specificity
This assay has high sensitivity and excellent specificity for detection of Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF).
No significant cross-reactivity or interference between Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) and analogues was observed.
Recovery
粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒Matrices listed below were spiked with certain level of recombinant Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) and the recovery rates were calculated by comparing the measured value to the expected amount of Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) in samples.
Matrix |
Recovery range (%) |
Average(%) |
serum(n=5) |
80-104 |
90 |
EDTA plasma(n=5) |
79-101 |
95 |
heparin plasma(n=5) |
96-105 |
99 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) 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 Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) 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) |
81-90% |
97-105% |
83-103% |
94-103% |
EDTA plasma(n=5) |
80-102% |
87-95% |
90-99% |
98-105% |
heparin plasma(n=5) |
85-95% |
86-97% |
79-96% |
90-99% |
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 immediately.
Test principle
粒细胞巨噬细胞集落刺激因子(GMCSF)检测试剂盒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 Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF). 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 Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF), 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 Colony Stimulating Factor 2, Granulocyte Macrophage (GMCSF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.
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