AffinityImmuno · ELISA Kit
Tocilizumab (Actemra®) Pharmacokinetic ELISA
Cat.No
EL-1611-202
1X96 wells
- Reactivity
- Human, mouse, rat
- Detection range
- 50ng/ml - 1.56ng/ml
- Sample type
- Serum Plasma
- Assay time
- 2.5 hours
- Antigen
- Tocilizumab (Actemra)
- Detection minimum
- 1.5ng/ml
Product Overview
The Tocilizumab ELISA kit is designed to measure free Tocilizumab with high specificity and sensitivity. Tocilizumab (Actemra®) is an immunosuppressive humanized monoclonal antibody drug, mainly for the treatment of rheumatoid arthritis (RA) and systemic juvenile idiopathic arthritis, a severe form of arthritis in children.
INTRODUCTION
Tocilizumab (Actemra®) is an immunosuppressive humanized monoclonal antibody drug, mainly for the treatment of rheumatoid arthritis (RA) and systemic juvenile idiopathic arthritis, a severe form of arthritis in children. Tocilizumab binds the interleukin-6 receptor (IL-6R) thus blocking the signaling caused by interleukin 6 (IL-6). IL-6 is a cytokine that plays an important role in immune response and is implicated in the pathogenesis of many diseases.
The Tocilizumab ELISA kit is designed to measure free Tocilizumab with high specificity and sensitivity.
PRINCIPLE OF THE ASSAY
This assay employs the sandwich enzyme immunoassay technique. Anti- Tocilizumab is coated onto a 96 well microplate. Calibrator and test samples are pipetted into the appropriate wells. Tocilizumab present in biological matrices is bound by the immobilized anti- Tocilizumab antibody. After washing away any unbound substances, enzyme linked anti- Tocilizumab antibody is added to the wells. This antibody is developed and purified specifically against Actemra® (domain residing in Fc portion of the Actemra® 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 Tocilizumab present in test samples. The color development is stopped and the intensity of the color is measured.
Target background
Tocilizumab (Actemra®) is an immunosuppressive humanized monoclonal antibody drug, mainly for the treatment of rheumatoid arthritis (RA) and systemic juvenile idiopathic arthritis, a severe form of arthritis in children. Tocilizumab binds the interleukin-6 receptor (IL-6R) thus blocking the signaling caused by interleukin 6 (IL-6). IL-6 is a cytokine that plays an important role in immune response and is implicated in the pathogenesis of many diseases.
Specifications
- Gene ID
- 3570
- Reactivity
- Human, mouse, rat
- Detection range
- 50ng/ml - 1.56ng/ml
- Sample type
- Serum Plasma
- Sample volume
- 15ul
- Assay time
- 2.5 hours
- Assay precision
- <10%, <10%
- Storage
- -20C, 1 year
- Size
- 1X96 wells
- Antigen
- Tocilizumab (Actemra)
- Detection minimum
- 1.5ng/ml
- Method
- Direct sandwich ELISA
- Detection method
- Peroxidase / OD450
- Application
- Quantification of Tocilizumab in biological matrices
- Plate format
- Strip
- Specificity
- Tocilizumab
- Preservative
- None
Kit Components
- 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
Assay Information
Reagent preparation
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 1/10 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 2,500 ng/mL to 78 ng/mL. The following is an example calibrator curve.
Results calculation
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 calibrator diluent and retested. If the samples have been diluted, the concentration determined from the standard curve must be multiplied by the dilution factor.
Sample collection
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.
Sample preparation
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.