BMDMs were plated in 12-well dishes at a density of 106 cells per well and infected with an MOI of 10. One hour postinfection the cells were washed and media containing gentamicin (50 μg ml−1) was added. Where indicated, 2 mM buthionine sulfoximine (Santa Cruz Biotechnology) was added to cells 16 h before infection and included throughout the infection. Four hours post-infection the cells were washed and harvested in LDS buffer containing 5% BME. The samples were then boiled and separated by 10% SDS–PAGE. A rabbit polyclonal antibody against the N terminus of ActA30 (link) and a mouse monoclonal antibody against P60 (Adipogen) were each used at a dilution of 1:5,000. P60 is a constitutively expressed bacterial protein31 (link) used as a loading control.
Buthionine sulfoximine
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Buthionine sulfoximine is a synthetic amino acid that functions as an inhibitor of glutathione synthesis. It acts by competitively inhibiting the enzyme gamma-glutamylcysteine synthetase, which is a key step in the production of glutathione.
Automatically generated - may contain errors
Lab products found in correlation
Mouse monoclonal antibody against p60, by Adipogen (1 mentions)
Proteinase inhibitor, by Roche (1 mentions)
Mntbap chloride, by Santa Cruz Biotechnology (1 mentions)
M2128, by Merck Group (1 mentions)
Glutathione monoethyl ester, by Santa Cruz Biotechnology (1 mentions)
Dcf d6883, by Merck Group (1 mentions)
Cabozantinib, by MedChemExpress (1 mentions)
Hoechst 33258 b 1155, by Merck Group (1 mentions)
A 1331852, by MedChemExpress (1 mentions)
Aβ1 42 peptide, by Bachem (1 mentions)
3 protocols using buthionine sulfoximine
1
Quantifying Listeria ActA Expression
2
Cytotoxicity Evaluation of Targeted Therapeutics
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Dulbecco’s Modified Eagle’s Medium (DMEM), trypsin-EDTA, penicillin-streptomycin and dimethyl sulfoxide (DMSO), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (M2128), Hoechst 33258 (B1155) and DCF (D6883) were purchased from Sigma–Aldrich (St. Louis, MO, USA). All tissue culture-ware was from Nunc (Roskilde, Denmark). Proteinase inhibitors were from Roche (Madrid, Spain). ECL western blotting substrate was from Pierce (Thermo Fisher Scientific, Rockford, IL, USA). Novex Sharp Pre-Stained Protein Standard (LC5800) (T-3168) were from Invitrogen Life Technologies (Carlsbad, CA, USA). Sorafenib (BAY 43-9006, Nexavar) and Regorafenib (BAY 73-4506, Stivarga) are manufactured by Bayer. Cabozantinib and A-1331852 were purchased from MedChem Express (Monmouth Junction, NJ, USA). Buthionine sulfoximine, MnTBAP chloride and glutathione monoethyl ester were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).
3
Amyloid-beta Peptide Study Protocol
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Aβ1-42 peptide was purchased from Bachem (Bubendorf, Switzerland). Buthionine sulfoximine and iron sulfate heptahydrate were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Sigma (St. Louis, MO, USA), respectively. Rabbit polyclonal antiserum to Iba1 was ordered from Wako Chemicals GmbH, (Neuss, Germany). Monoclonal anti-Aβ antibody was obtained from Abcam (Cambridge, UK). Artificial CSF and MCC950 (Coll et al., 2015) (link) were purchased from Tocris (Bristol, UK) and Sigma, respectively.
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