Bevacizumab (Avastin) Pharmacokinetic ELISA

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 It had been approved for breast cancer, but that approval was withdrawn when later studies showed no evidence of effectiveness

Product Specifications

CAS Number

7732-18-5

Gene ID

7422

Reactivity

Human, mouse, rat

Reaction Volume

15 µL

Target Antigen

Bevacizumab (Avastin)

Detection Method

Peroxidase / OD450

Assay Type

ELISA

Assay Principle

Quantification of Bevacizumab in biological matrices

Assay 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.

Assay Performance Time

2.5 hours

Sample Type

Serum Plasma

Sample Volume

15ul

Detection Range

50ng/ml - 1.56ng/ml

Precision

<10%, <10%

Sensitivity

Bevacizumab

Limit Of Detection

1.5ng/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

Shipping Conditions

Blue Ice

Storage Conditions

-20C, 1 year

Shelf Life

12 months

Badge

Hot

Specificity

Bevacizumab

Method

Direct sandwich ELISA

Preservative

None

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.

Operating 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.

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.

Plate

Strip

Curated Selection

Explore Other Products

Discover premium biology products from our extensive collection of 20M+ items

T0081 1x106 cells / 1.0 mL

Immortalized Fetal Bovine Kidney Cells - SV40

Ask

View Details

MBS9920658 Inquire

Donkey FAT Tumor Suppressor Homolog 1 (FAT1) ELISA Kit

Ask

View Details