Bevacizumab

Catalog No.A2006 Synonyms: rhuMab VEGF Non-humanized mouse model applicable

For research use only. Not for use in humans.

Bevacizumab is a humanized anti-VEGF monoclonal antibody which binds to and neutralizes all human VEGF-A isoforms and bioactive proteolytic fragments, MW:149 KD.

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Biological Activity

Description Bevacizumab is a humanized anti-VEGF monoclonal antibody which binds to and neutralizes all human VEGF-A isoforms and bioactive proteolytic fragments, MW:149 KD.
Targets
VEGF [1]
()
In vitro

Gly88 in human VEGF is essential for binding bevacizumab and this residue also underlies the species specificity of bevacizumab binding, since a serine residue is found in mouse and rat VEGF at the corresponding position. In common with its mouse counterpart, bevacizumab binds to and neutralizes all human VEGF-A isoforms and bioactive proteolytic fragments. It does not neutralize other members of the VEGF gene family, such as VEGF-B or VEGF-C[1].

In vivo The terminal half-life of bevacizumab in humans is 17-21 days. Bevacizumab inhibits the growth of human tumour cell lines in nude mice, achieving a maximal inhibition at the dose of 1-2 mg per kg twice weekly. Half-maximal inhibition requires 0.1-0.5 mg per kg doses. Safety evaluation studies of bevacizumab are conducted in Macaca fascicularis (cynomolgus monkey), a species in which bevacizumab is expected to be pharmacologically active, considering the complete identity between human and cynomolgus VEGF isoforms at the protein level. Following administration of bevacizumab for four or thirteen weeks, young adult cynomolgus monkeys exhibits a physeal dysplasia characterized by a dose-related increase in hypertrophied chondrocytes and inhibition of vascular invasion of the growth plate, which is very similar to the growth-plate lesion observed in mice treated with Flt(1-3)-IgG. Other expected effects of prolonged bevacizumab administration are suppression of angiogenesis in the female reproductive tract, resulting in decreased ovarian and uterine weights, and an absence of corpora lutea. Both the growth-plate and ovarian changes are reversible with cessation of treatment. Importantly, no other treatment-related effects are observed following bevacizumab administration at doses up to 50 mg per kg[1]. After s.c. administration, rhuMAb VEGF has a bioavailability of 69% in rats and 100% in mice and cynomolgus monkeys. It binds to primate VEGF and with lower affinity to rabbit VEGF but not to rat or mouse VEGF[2].

Protocol

Cell Research:
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  • Objective: Effects of bevacizumab on VEGF-induced HUVEC proliferation
    Cells: Human umbilical vein endothelial cells (HUVECs)
    Concentrations: 0~500 ng/mL
    Incubation Time: 96 h
    Method: 50 ng/ml of rhVEGF was pre-incubated with a concentration range of bevacizumab (0-500 ng/ml) for 2 h before added to HUVACs. The plate was continuously incubated for 4 days. The proliferation of cells was determined by using Alamar Blue dye.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/15886877

    Objective: Cell proliferation assay
    Cells: non small cell lung cancer (NSCLC), colorectal cancer (CRC), breast cancer (BC) and renal cell carcinoma (RCC)
    Concentrations: 250 μg/ml
    Incubation Time: 24, 48, 72 or 96 hours
    Method: Between 2 × 103 and 5 × 103 cells/well (cell line/doubling time dependent) are seeded into 96 well plates and incubated overnight to adhere. Medium is then replaced by RPMI-1640 with reduced FBS and bevacizumab or VEGFA at the concentrations indicated (time point zero). After 24, 48, 72 or 96 hours in hypoxia, MTT (5 mg/ml in PBS) is added and incubated for 2 hours at 37°C. The supernatant is removed and reaction products are solubilized for 1 h in 10% HCl, 0.1% NP-40 in isopropanol. Absorbance is measured at 570 nm with a reference wavelength of 650 nm using an ELISA reader. Each experimental condition is analyzed in triplicate and results are an average of a minimum of three biological repetitions.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/24059699

    Bevacizumab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)
Animal Research:
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  • Objective: To determine the antitumor response of bevacizumab and cisplatin as single agents or in combination in xenograft models of ovarian cancer
    Animal Models: Nude mice were injected (i.p.) with 5×106 A2780 cells
    Formulation: PBS
    Dosages: 5mg/kg
    Administration: i.p.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/19047105

    Objective: To determine the pharmacokinetics of bevacizumab
    Animal Models: Female nude mice
    Formulation: --
    Dosages: 9.3 mg/kg
    Administration: i.v. or s.c. injection
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/9862791

    Bevacizumab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)

Product Details

Formulation PBS buffer, pH 7.2
Isotype Human IgG
Source CHO cells
Storage Store at -80°C and avoid freeze-thaw cycles.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID