Anti-PEG antibody ELISA Kit (mouse IgM specific)

适用物种:mouse

样本类型:Serum,Plasma

检测范围:62.5 ng/mL -1000 ng/mL

最低检出限:62.5ng/ml

测定时间:2.5 hours

检测格式:Direct sandwich ELISA

检测原理:
This assay employs the sandwich enzyme immunoassay technique. Immobilized PEG is coated onto a 96 well microplate. Calibrator and test samples are pipetted into the appropriate wells. Anti-PEG antibodies present in biological matrices is bound by the immobilized PEG. After washing away any unbound substances, enzyme linked anti IgG or IgM antibody 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 anti-PEG antibodies 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:Anti-PEG mouse IgM antibodies

应用:Quanitification of mouse IgM specific

包被板类型:Strip

样品体积:15ul

特异性:Anti-PEG antibodies

检测精度(批内差异,批间差异):<10%, <10%

防腐剂:None

 试剂盒组分:
Coated microtiter plate, 96 wells
QC samples - 6x250ul
10X wash buffer - 50 ml
Assay buffer - 50ml
1000X detection reagent - 17ul
TMB - 12ml
TMB stop solution - 12ml
Plate sealers - 3

储存温度和稳定性:Stable at -20°C for 1 year

背景描述:
Polyethylene glycol (PEG) chains are often used to modify therapeutic biologic agents in order to prolong the circulating half-life of the modified protein. It has been reported that repeat injections of PEGylated proteins can induce anti-PEG antibodies.

检测程序:
This assay employs the sandwich enzyme immunoassay technique. Immobilized PEG is coated onto a 96 well microplate. Calibrator and test samples are pipetted into the appropriate wells. Anti-PEG antibodies present in biological matrices is bound by the immobilized PEG. After washing away any unbound substances, enzyme linked anti IgG or IgM antibody 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 anti-PEG antibodies 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 test samples 1/5 with assay buffer before use (for example add 50μl of test sample to 200μl assay buffer). Mix well.

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 deionized water 1/10 before use (for example add 20mL concentrate to 180mL deionized 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.

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. Alternatively a log-log curve fit may be used.
2. The concentration of the unknowns can be read directly from this standard curve using the absorbance value for each sample.
3. Any sample undiluted or diluted still reading greater than the highest standard should be diluted appropriately with assay buffer and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.



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