Rituximab (Rituxan) Pharmacokinetic ELISA Kit
适用物种:Human,mouse,rat

样本类型:Serum,Plasma

检测范围:100ng/ml - 5ng/ml

最低检出限:5ng/ml

测定时间:2.5 hours

检测格式:Direct sandwich ELISA

检测原理:
The Rituximab ELISA kit is designed to measure free Rituximab with high specificity and sensitivity . This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Rituximab 96 well plate is provided. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Rituximab present in biological matrices is bound by the immobilized capture antibody. After washing away any unbound substances, enzyme linked detection 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 Rituximab present in test samples and the concentration is calculated from the standard series.

检测方法:Peroxidase / OD450 

靶抗原:Rituximab (Rituxan)

应用:Quantification of Rituximab in biological matrices

包被板类型:Strip

样品体积:15ul

特异性:Rituximab

检测精度(批内差异,批间差异):<10%, <10%

防腐剂:None

Gene ID:931

试剂盒组分:
Coated microtiter plate, 96 wells
Calibrator diluent. - 1.8ml
Calibrator 12ul
10X wash buffer - 25ml
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:
Rituximab is a chimeric monoclonal antibody used in the treatment of diseases characterized by abnormal or excessive B cells. Rituximab targets CD20, which is expressed on the surface of B cells, to induce apoptosis in B cells and is used in the treatment of leukemias and lymphomas, some autoimmune disorders, and organ transplant. A quantitative method for Rituximab has been developed and optimized for pharmacokinetic assessment of samples.

检测程序:
This assay employs the sandwich enzyme immunoassay technique. Anti- Rituximab is coated onto a 96 well microplate. Calibrator, quality control samples (if desired) and test samples are pipetted into the appropriate wells. Rituximab present in biological matrices is bound by the immobilized anti- Rituximab antibody. After washing away any unbound substances, enzyme linked anti- Rituximab antibody is added to the wells. This antibody is developed and purified specifically against truncated Rituxan® (domain residing in Fc portion of the Rituxan® molecule). 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 Rituximab 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. *Note that test samples may require further dilution when peak values are predicted to exceed 5ug/ml. Do not store diluted samples.

试剂制备:
Prepare 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 1/10 before use (for example add 20mL concentrate to 180mL 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. Preparation of Calibrators: Prepare calibrators with concentrations ranging from 5000 ng/mL to 250 ng/ mL.

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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