Bevacizumab ELISA Kit

Catalog No. ABIN4886390

Human, Mouse and Rat Bevacizumab ELISA Kit, Colorimetric assay for quantification of Human, Mouse and Rat Bevacizumab.

Quick Overview for Bevacizumab ELISA Kit (ABIN4886390)

Target: Bevacizumab

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 1.56 ng/mL - 50 ng/mL

Application: ELISA

Sample Type: Plasma, Serum

Product Details Bevacizumab ELISA Kit

Minimum Detection Limit: 1.56 ng/mL

Purpose: Quantification of Bevacizumab in biological matrices

Analytical Method: Quantitative

Specificity: Bevacizumab (Avastin)

Cross-Reactivity (Details): hIgG1, rituximab, and infliximab prepared at 250 ng/mL were assayed and exhibited no crossreactivity or interference.

Sensitivity: 1.5 ng/mL

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

Material not included:

• Precision pipettes calibrated to deliver 5-1000μL
• Multi-channel pipette calibrated to deliver 50-200μL
• Plate shaker
• Disposable tips
• Vortex-Mixe
• Distilled or de-ionized water
• Microplate reader capable of reading 450nm with background subtraction

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 15 μL

Assay Time: 2.5 h

Plate: Pre-coated

Protocol: The Bevacizumab ELISA kit is designed to measure free Bevacizumab with high specificity and sensitivity. This assay employs the sandwich enzyme immunoassay technique. A precoated anti-Bevacizumab 96 well plate is provided. 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 development is proportional to the amount of Bevaciuzumab present in test samples and the concentration is calculated from the standard series.

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 20 mL concentrate to 180 mL ultra-pure water). Mix well. 2. Detection Reagent (1X) Preparation: Dilute detection reagent with assay buffer 1/1000 before use (for example add 11 μL concentrate to 11 mL 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.

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.

Assay Procedure:

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 development is proportional to the amount of Bevacizumab present in test samples.

Calculation of Results:

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.

Assay Precision: Intra-assay precision: <10%
Inter-assay precision: <10%

Restrictions: For Research Use only

Handling

Preservative: Without preservative

Storage: -20 °C

Storage Comment: -20C, 1 year

Expiry Date: 12 months

Target Details for Bevacizumab

Target: Bevacizumab

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

Gene ID: 7422