AffinityImmuno · ELISA Kit
Bevacizumab (Avastin®) Pharmacokinetic ELISA Kit
Cat.No
EL-1611-021
1X96 wells
- Reactivity
- Human, mouse, rat
- Detection range
- 50ng/ml - 1.56ng/ml
- Sample type
- Serum Plasma
- Assay time
- 2.5 hours
- Antigen
- Bevacizumab (Avastin)
- Detection minimum
- 1.5ng/ml
Product Overview
The Bevacizumab ELISA kit is designed to measure free Bevacizumab with high specificity and sensitivity, through use of a pair of antibodies.
INTRODUCTION
The Bevacizumab ELISA kit is designed to measure free Bevacizumab with high specificity and sensitivity. The assay design uses a pair of antibodies allowing detection of whole Bevacizumab molecules in biological matrices. Bevacizumab (Trade name Avastin®) is a recombinant humanized monoclonal antibody that blocks angiogenesis by inhibiting vascular endothelial growth factor A(VEGF-A). VEGF-A is a chemical signal that stimulates angiogenesis in a variety of diseases, especially in cancer. Bevacizumab was the first clinically available angiogenesis inhibitor in the United States.
PRINCIPLE OF THE ASSAY
This assay employs the sandwich enzyme immunoassay technique. Anti-Bevacizumab is coated onto a 96 well microplate. Calibrator, quality control samples and test samples are pipetted into the appropriate wells. Bevacizumab 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 devel opment is proportional to the amount of Bevaciuzumab present in test samples.
Target background
Bevacizumab (Trade name Avastin®) is a recombinant humanized monoclonal antibody that blocks angiogenesis by inhibiting vascular endothelial growth factor A (VEGF-A). VEGF-A stimulates angiogenesis in a variety of diseases, including cancer. Bevacizumab was the first clinically available angiogenesis inhibitor in the United States. Bevacizumab was approved by the U.S. Food and Drug Administration (FDA) for certain metastatic cancers. It received its first approval in 2004, for combined use with standard chemotherapy for metastatic colon cancer. It has since been approved for use in certain lung cancers, renal cancers, ovarian cancers, and glioblastoma multiforme of the brain. It had been approved for breast cancer, but that approval was withdrawn when later studies showed no evidence of effectiveness
Specifications
- Gene ID
- 7422
- 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
- Bevacizumab (Avastin)
- Detection minimum
- 1.5ng/ml
- Method
- Direct sandwich ELISA
- Detection method
- Peroxidase / OD450
- Application
- Quantification of Bevacizumab in biological matrices
- Plate format
- Strip
- Specificity
- Bevacizumab
- 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.