MDA-MB-231, MDA-MB-468, MDA-MB-453 and MDA-MB-361 breast cancer cells lines were grown in SILAC-DMEM (Euroclone) supplemented with 2 mM l -glutamine, 146 mg/l of lysine (Sigma-Aldrich), 84 mg/l l -13C615N4-arginine (Arg-10, Sigma-Aldrich), 10% dialyzed serum (Life Technologies) and penicillin/streptomycin for at least eight doublings to obtain complete labeling with heavy-labeled aminoacids. Histones were isolated as previously described (30 (link)), mixed in equal amounts, lyophilized, and stored at −80°C until use. Because the most abundant histone PTMs found in GBM and ovarian cancer samples are present in the breast cancer super-SILAC mix, the same standard was used to analyze all samples.
L 13c615n4 arginine arg 10
Manufactured by Merck Group
L-13C615N4-arginine (Arg-10) is a stable isotope-labeled amino acid product. It is a form of the amino acid arginine labeled with the stable isotopes carbon-13 and nitrogen-15. This product is intended for use in research applications that require the incorporation of stable isotope-labeled amino acids.
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Lab products found in correlation
2 protocols using l 13c615n4 arginine arg 10
1
Breast Cancer Cell Line SILAC-Labeled Histones
2
Histone-Focused SILAC Labeling Approach
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A histone-focused super-stable isotope labeling by amino acid in cell culture (SILAC) approach was used as we have recently described [12 (link)]. MDA-MB-231, MDA-MB-468, MDA-MB-453, and MDA-MB-361 breast cancer cells lines were grown in SILAC-DMEM (Euroclone) supplemented with 2 mM l -glutamine, 146 mg/l of lysine (Sigma-Aldrich), 84 mg/l l -13C615N4-arginine (Arg-10, Sigma-Aldrich), 10% dialyzed serum (Life Technologies), and penicillin/streptomycin for at least eight doublings to obtain complete labeling with heavy-labeled aminoacids. Histones were isolated from the different cell lines through nuclei isolation on a sucrose cushion followed by acidic extraction, as described [13 (link)], mixed in equal amounts, lyophilized, and stored at −80 °C until use.
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