Synthesis and Characterization of Maytansinoid Compounds

S-methyl DM1 and other maytansinoids were synthesized as described previously [6 (link)]. ³[H]-S-methyl DM1 was synthesized at American Radiolabeled Chemicals. 1,2-dioleoyl-sn-glycero-3-phosphocholine in chloroform (Avanti Polar Lipids) was stored under argon at-20^o C. ³[H]-Paclitaxel was from Moravek Biochemicals. MCF7 cell tubulin and bovine brain tubulin (>99%; isolated by the method described in [10 (link)]) were purchased from Cytoskeleton, Inc. Paclitaxel was from Tocris; nocodazole, vinblastine sulfate, demecolcine, monoclonal anti-α-tubulin clone B-5–1–2 murine IgG1 T6074, RIPA buffer from Sigma; Halt Protease & Phospatase Inhibitor Cocktail (no EDTA or other chelators) was from Thermo Scientific; DilC18(3) and MagicMark XP protein size marker were purchased from Invitrogen; peroxidase-conjugated AffiniPure F(ab’)2 fragment goat anti mouse IgG (H+L) 115–036–062 was purchased from Jackson Labs; ECL Advanced Blocking Agent CPK1075 was purchased from GE Healthcare.

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Goldmacher V.S., Audette C.A., Guan Y., Sidhom E.H., Shah J.V., Whiteman K.R, & Kovtun Y.V. (2015). High-Affinity Accumulation of a Maytansinoid in Cells via Weak Tubulin Interaction. PLoS ONE, 10(2), e0117523.

Publication 2015

3[H]-paclitaxel bovine brain tubulin Paclitaxel nocodazole vinblastine sulfate demecolcine peroxidase-conjugated AffiniPure F(ab’)2 fragment goat anti mouse IgG (H+L)

Agent Anti igg Argon Bovine Brain Buffer Chelators Chloroform Clone Cytoskeleton Demecolcine Edta Goat Igg1 Lipids Mcf7 cell Mouse Nocodazole Paclitaxel Peroxidase Protease inhibitor Protein Ripa Sn glycero 3 phosphocholine Tubulin Vinblastine sulfate α tubulin

Corresponding Organization : ImmunoGen (United States)

Other organizations : Center for Systems Biology, Harvard University, Brigham and Women's Hospital

Top 5 similar protocols

Variable analysis

independent variables

Synthesis of S-methyl DM1 and other maytansinoids
Synthesis of ³[H]-S-methyl DM1

dependent variables

Measurement of ³[H]-S-methyl DM1
Measurement of ³[H]-paclitaxel

control variables

Storage of 1,2-dioleoyl-sn-glycero-3-phosphocholine in chloroform under argon at -20°C
Isolation of MCF7 cell tubulin and bovine brain tubulin
Use of paclitaxel, nocodazole, vinblastine sulfate, and demecolcine
Use of monoclonal anti-α-tubulin clone B-5–1–2 murine IgG1 T6074
Use of RIPA buffer, Halt Protease & Phospatase Inhibitor Cocktail, DilC18(3), MagicMark XP protein size marker, peroxidase-conjugated AffiniPure F(ab')2 fragment goat anti mouse IgG (H+L) 115–036–062, and ECL Advanced Blocking Agent CPK1075

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