Trastuzumab

Catalog No.A2007 Synonyms: Herceptin Non-humanized mouse model applicable

For research use only. Not for use in humans.

Trastuzumab is a humanized, recombinant monoclonal antibody that binds to the extracellular domain of HER2, MW:145.53 KD.

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Biological Activity

Description Trastuzumab is a humanized, recombinant monoclonal antibody that binds to the extracellular domain of HER2, MW:145.53 KD.
Targets
HER2 [1]
()
In vitro

Trastuzumab has been demonstrated to exert a variety of anti-tumor effects selectively in HER2-overexpressing tumor cells. Trastuzumab binds to the juxtamembrane domain of HER2 and upon receptor binding, the antibody downregulates the expression of HER2. Trastuzumab selectively blocks ligand-independent HER2-HER3 dimerization. In addition, trastuzumab binding to HER2 blocks proteolytic cleavage of the extracellular domain of HER2, resulting in diminished levels of the more active p95-HER2 form of HER2. As a result of these effects on the HER2 receptor, trastuzumab causes downregulation of PI3K pathway signaling and downstream mediators of cell cycle progression such as cyclin D1. Trastuzumab not only inhibits HER2 signaling pathways but also triggers immune-mediated responses against HER2-overexpressing cells. Trastuzumab binding engages Fc receptors on immune effector cells leading to antibody-dependent cellular cytotoxicity. Beyond these effects, trastuzumab has been shown to have antiangiogenic effects and to lower the proapoptotic threshold for chemotherapy[1].

In vivo Trastuzumab is shown to be safe when administered chronically to a range of animals including primates[1]. In experiments mimicking adjuvant therapy of submacroscopic disease in vivo (JIMT-1 cells inoculated s.c. in severe combined immunodeficiency mice), trastuzumab is able to inhibit the outgrowth of macroscopically detectable xenograft tumors for up to 5-7 weeks. Trastuzumab has a remarkable antitumor effect and is currently used worldwide for the treatment of breast cancer. In in vivo models, trastuzumab decreases the microvessel density of breast cancer xenografts. It is well documented that trastuzumab acts directly in cancer cell signaling, as well as indirectly via the immune system[2].

Protocol

Cell Research:
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  • Objective: In vitro Assay of Drug Sensitivity
    Cells: JIMT-1, SKBR-3, and BT-474 cells
    Concentrations: 0, 1, 10, or 100 μg/mL
    Incubation Time: 72 h
    Method: Exponentially growing cells are harvested and plated in single wells of a 96-well flat-bottomed tissue culture plate at defined densities, ranging from 4,500-8,000 cells per well depending on the cell line. After overnight culture, the regular medium is exchanged to medium containing 0, 1, 10, or 100 μg/mL trastuzumab or trastuzumab-F(ab′)2. Cell viability is tested after 72 h of treatment. Fluorescence is detected at an excitation of 544 nm, and emission is detected at 590 nm.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/17620435

    Objective: Antibody-dependent cellular cytotoxicity (ADCC)
    Cells: Peripheral blood lymphocytes (PBLs), USPC cell lines (Uterine cancer cell)
    Concentrations: 2.5 μg/mL
    Incubation Time: --
    Method: A standard 5 h chromium (51Cr) release assay was performed to measure the cytotoxic reactivity of peripheral blood lymphocytes (PBLs) from several healthy donors in combination with trastuzumab and/or pertuzumab against USPC target cell lines.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/19920829

    Ttrastuzumab can apply to immunodeficient mice (eg: SCID mice), various cancer cell lines and other related assays (Only for Reference)
Animal Research:
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  • Objective: Xenograft Tumor Assays
    Animal Models: SCID C.B-17 scid/scid and nu/nmri nude mouse
    Formulation: saline
    Dosages: 5 μg/g
    Administration: i.p.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/17620435

    Objective: To investigate the antitumor activity of the combination of pertuzumab and trastuzumab for gastric cancer
    Animal Models: BALB-nu/nu mice were injected (s.c.) with 5 × 106 Calu-3 cells
    Formulation: --
    Dosages: 30 mg/kg loading dose, then 15 mg/kg weekly
    Administration: i.p.
    Reference: https://www.ncbi.nlm.nih.gov/pubmed/19934333

    Ttrastuzumab can apply to immunodeficient mice (eg: SCID 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