Filgrastim (GCSF) Pharmacokinetic ELISA Kit

适用物种:human

样本类型:Serum,Plasma

检测范围:9.375ng/mL - 500 ng/mL

最低检出限:9.375 ng/mL

测定时间:2.5 hours

检测格式:Direct sandwich ELISA

检测原理:
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.

检测方法:Peroxidase / OD450 

靶抗原:granulocyte colony stimulating factor (G-CSF)

应用:granulocyte colony stimulating factor in biological matrices

包被板类型:Strip

样品体积:15ul

特异性:GCSF

检测精度(批内差异,批间差异):< 25% for ULOQ and LLOQ and<20% for the remaining concentrations

防腐剂:None

Gene ID:1440

试剂盒组分:
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 17μl
1000X detection reagent 17μl
TMB 12ml
TMB stop solution 12ml

储存温度和稳定性:
Stable at -20°C for 1 year

背景描述:
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

检测程序:
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.

样本收集:
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.

样品制备:
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.

试剂制备:
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 25mL concentrate to 475mL ultrapure water). Mix well.
2. Secondary Antibody (1X) Preparation: Dilute secondary antibody with assay buffer 1/1000 before use (for example add 12μl to 12mL of assay buffer). Mix well.
3. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 12μl concentrate to 12ml of assay buffer). Mix well.
4. Calibrator Preparation: Dilute the calibrator from 1mg/ml down to 5µg/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.

结果计算:
1. 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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