Slicing and culturing of 300 μm thick heart tissue slices were performed as previously described31 (link), 38 . A refined oxygenated growth medium was used (Medium 199, 1x ITS supplement, 10% FBS, 5 ng/mL VEGF, 10 ng/mL FGF-basic, and 2x Antibiotic-Antimycotic), and changed 3 times/day. Sunitinib (Tocris Inc.) (100 nM, 1μM, and 10 μM), trastuzumab (InvivoGen Inc.) (1 μg, 10 μg, and 100 μg), or doxorubicin (Sigma Millipore Inc.) (100 nM, 1μM, and 10 μM, and 50 μM) was added freshly to the culture medium at each medium change. Control slices received DMSO at the same dilution factor as the matched drug-treated slices. For detailed protocols regarding slicing and culturing heart slices, please see the Supplementary Methods .
Trastuzumab
Manufactured by InvivoGen
Trastuzumab is a monoclonal antibody that binds to the HER2/neu (human epidermal growth factor receptor 2) receptor on the surface of certain cancer cells. It is used in the detection and analysis of HER2 expression in various research applications.
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Lab products found in correlation
3 protocols using trastuzumab
1
Culturing Thick Heart Tissue Slices
2
Antibody Characterization for Immunotherapy
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Trastuzumab and Rituximab antibodies were purchased from Invivogen: anti-HER2-Tra-hIgG1 (her2tra-mab1), anti-hCD20-hIgG1 (hcd20-mab1), anti-hCD20-hIgG1NQ (hcd20-mab12), anti-hCD20-hIgG2 (hcd20-mab2), anti-hCD20-hIgG3 (hcd20-mab3), anti-hCD20-IgG4 (hcd20-mab4), anti-hCD20-mIgG1 (hcd20-mab9), and anti-CD20-mIgG2a (hcd20-mab10). mIgG2b isotype control clone MPC-11 (559530) was purchased from BD. CR9114, FI6, 2B06, 2G02, 6F12, 8H9, and KB2 were kindly provided by Florian Krammer; see also Table 1 .
3
Recombinant IgG2 Trastuzumab Production
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The clinically approved antibodies cetuximab (chimeric human IgG1; clone 225, Erbitux®), panitumumab (human IgG2; clone E7.6.3, Vectibix®) and trastuzumab (humanized IgG1; clone 4D5-8; Herceptin®) were obtained from Merck (Darmstadt, DE), Amgen (Thousand Oaks, California, USA) and Roche (Basel, CH), respectively. The recombinant IgG2 variant of trastuzumab was expressed transiently from the pFUSE vector system (Invivogen) in Expi293 cells (Thermo Fisher). Pure monomeric fractions were obtained by affinity purification (CaptureSelect CH1) (Thermo Fisher) and size exclusion chromatography (SEC) (Superdex 200 Increase) (Cytiva Lifesciences) using an ÄKTA Avant 25 instrument.
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