GCSF ADA (granulocyte colony stimulating factor) ELISA Kit

The AffinityImmuno GCSF ADA ELISA kit contains the components required for measuring antibodies to GCSF in biological matrices such as human serum, plasma or cell culture supernatant.

适用物种:Human

样本类型:Serum,Plasma

检测范围:45 ng/mL -1440 ng/mL

最低检出限:45 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 enzyme immunoassay technique. GCSF is coated onto a 96 well microplate. Calibrator and test samples are prepared by dilution into assay buffer and are pipetted into the appropriate wells. Antibodies to GCSF present in biological matrices are bound to the immobilized GCSF. After washing away any unbound substances, antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of antibodies to GCSF present in the calibrator and test samples. The color development is stopped and the intensity of the color is measured.

检测方法:Peroxidase / OD450 

靶抗原:antibodies to GCSF

应用:Quantification of antibodies to G-CSF

包被板类型:Strip

样品体积:15ul

特异性:anti-GCSF antibodies

检测精度(批内差异,批间差异):<20%, <20%

防腐剂:None

试剂盒组分:
Coated microtiter plate, 96 wells (12x8 strips) 1
Calibrator diluent 1.8 ml
Calibrator (1.15mg/mL) 12 μl
10X wash buffer 50 ml
Assay buffer 50 ml
1000X detection reagent 17 μl
TMB 12 ml
TMB stop solution

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 enzyme immunoassay technique. GCSF is coated onto a 96 well microplate. Calibrator and test samples are prepared by dilution into assay buffer and are pipetted into the appropriate wells. Antibodies to GCSF present in biological matrices are bound to the immobilized GCSF. After washing away any unbound substances, antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of antibodies to GCSF present in the calibrator and 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 50mL wash buffer concentrate to 450mL ultra-pure water). Mix well.
2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11μl concentrate to 11ml of assay buffer). Mix well.
3. Calibrator Preparation: Dilute the calibrator from 1.15mg/ml down to 11.5µg/ml by pipetting 5µl of calibrator stock into 495µl assay buffer. Label "Cal. Int." Mix well. Prepare calibrators with concentrations ranging from 1440ng/ml to 45ng/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.



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