G-CSF ELISA 试剂盒
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Quick Overview for G-CSF ELISA 试剂盒 (ABIN8118914)
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See all G-CSF (CSF3) ELISA试剂盒适用
检测方法
实验类型
检测范围
应用范围
样品类型
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最低检测浓度
- 9.375 ng/mL
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原理
- granulocyte colony stimulating factor in biological matrices
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Analytical Method
- Quantitative
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特异性
- GCSF
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组件
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- Coated microtiter plate, 96 wells
- (1x8 strips) 1 Calibrator diluent 1.8ml
- Calibrator (1mg/ml) 12μl 20X wash buffer 25ml
- Assay buffer 50ml
- 1000X secondary antibody 1000X detection reagent 17μl TMB 12ml
- TMB stop solution 12ml
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试剂未包括
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- Precision pipettes calibrated to deliver 5-1000μL
- Multi-channel pipette calibrated to deliver 50-200μL
- Plate shaker
- Disposable tips
- Vortex-Mixe
- Distilled or de-ionized water
- Microplate reader capable of reading 450nm with background subtraction
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应用备注
- Optimal working dilution should be determined by the investigator.
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样本量
- 15 μL
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实验时间
- 2.5 h
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板类型
- Pre-coated
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实验流程
- This assay employs the indirect sandwich enzyme immunoassay technique. Anti-GCSF is coated onto a 96 well microplate. Calibrator and test samples prepared by dilution into assay buffer and are pipetted into the appropriate wells. GCSF present in biological matrices is bound by the immobilized anti- GCSF antibody. After washing away any unbound substances, tagged anti-GCSF secondary antibody is added to the wells. After washing, diluted detection reagent is added to the wells. The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of GCSF present in test samples. The color development is stopped and the intensity of the color is measured.
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试剂准备
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Prepare only the appropriate amount of required reagent on the day of use. Store all reagents as per instructions stated on the label. 1. Wash Buffer (1X) Preparation: Dilute wash buffer concentrate with ultra-pure water 1/20 before use (for example add 25 mL concentrate to 475 mL ultrapure water). Mix well. 2. Secondary Antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for example add 12 μL to 12 mL of assay buffer). Mix well. 3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12 μL concentrate to 12 mL of assay buffer). Mix well. 4. Calibrator Preparation: Dilute the calibrator from 1 mg/mL down to 5 μg/mL by pipetting 5μL of calibrator stock into 995μL assay buffer. Label "Cal. Int." Mix well. Prepare calibrators with concentrations ranging from 500 ng/mL to 9.375 ng/mL. The following is an example calibrator curve.
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样品收集
- This kit is compatible with EDTA-plasma, heparinplasma and serum samples. Samples can be stored at or below -20 °C for up to 1 year.
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样品制备
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Dilute calibrators and test samples 1/50 with assay buffer (for example add 5μL of prepared calibrator or sample to 245μL of assay buffer). Mix well. Do not store diluted samples.
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结果分析
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- Construct a standard curve by plotting the absorbance obtained from each standard against concentration. Use a 4 or 5 parameter curve fit. 2. The concentration of the unknowns can be back calculated directly from this standard curve using the absorbance value for each sample. 3. Any sample diluted more or less than the standard series will need additional data correction. For example, if the sample is diluted 1/50, then the concentration will be calculated by dividing by 2 due to the calibrators being 2 times more diluted. Similarly, if the sample is diluted 1/500, then the concentration will be calculated by multiplying by a correction factor of 5 due to the calibrators being 5 times more concentrated.
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实验精密度
- < 25% for ULOQ and LLOQ and <20% for the remaining concentrations
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限制
- 仅限研究用
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储存液
- Without preservative
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储存条件
- -20 °C
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储存方法
- -20C, 1 year
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有效期
- 12 months
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- G-CSF (CSF3) (Colony Stimulating Factor 3 (Granulocyte) (CSF3))
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别名
- granulocyte colony stimulating factor
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背景
- Granulocyte colony-stimulating factor (G-CSF or GCSF), also known as colony-stimulating factor 3 (CSF 3), is a glycoprotein that stimulates the bone marrow to produce granulocytes and stem cells and release them into the bloodstream
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基因ID
- 1440
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途径
- Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization
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