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2 mercaptoethanol bme

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2-mercaptoethanol (BME) is a colorless liquid that is commonly used as a reducing agent in various laboratory applications. It functions by disrupting disulfide bonds, which are chemical linkages between sulfur atoms in proteins. This property makes BME useful in the preparation and analysis of biological samples.

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Lab products found in correlation

Fetal bovine serum (fbs), by Merck Group (3 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Laemmli sample buffer, by Bio-Rad (2 mentions) Glacial acetic acid, by Merck Group (1 mentions) Hplc grade chloroform, by Thermo Fisher Scientific (1 mentions) Optima lc ms grade methanol, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Formic acid, by Thermo Fisher Scientific (1 mentions) Acetonitrile, by Thermo Fisher Scientific (1 mentions) Isopropanol, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Ammonium acetate, by Thermo Fisher Scientific (1 mentions) Peroxide free arachidonic acid, by Cayman Chemical (1 mentions) Hca 7, by American Type Culture Collection (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions)

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2-mercaptoethanol (1795 citations) Mercaptoethanol (160 citations)

9 protocols using 2 mercaptoethanol bme

1

Synthesis and Analysis of Oxo-ETE Lipid Metabolites

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11-oxoeicosatetranoic acid (11-oxo-ETE), 15-oxoeicosatetranoic acid (15-oxo-ETE), 11-oxo-eicosatetranoic acid methyl ester (11-oxo-ETE-ME), 15-oxo-eicosatetranoic acid methyl ester (15-oxo-ETE-ME) and the [13C20]-labeled 15-oxo-ETE internal standard were synthesized in-house as previously reported (2 (link)). [13C315N1]-Arachidonoyl-CoA used as an internal standard was synthesized as previously reported (19 (link)). Peroxide-free arachidonic acid (AA) was purchased from Cayman Chemical (Ann Arbor, MI). 5-sulfosalicilic acid (SSA), ammonium formate, glacial acetic acid, 2-mercaptoethanol (BME), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO). HPLC grade chloroform as well as Optima LC-MS grade methanol, acetonitrile, water, isopropanol, ammonium acetate, and formic acid were purchased from Fisher Scientific (Pittsburgh, PA). 2-(2-pyridyl) ethyl functionalized silica gel (100 mg/mL) solid phase extraction (SPE) cartridges were obtained from Supelco Analytical (Bellefonte, PA). Streptomycin and penicillin were purchased from Invitrogen (Carlsbad, CA). Fetal bovine serum (FBS) was from Gemini Bioproducts (West Sacramento, CA). Human colorectal adenocarcinoma cells (LoVo and HCA-7) were obtained from ATCC (Manassas, VA).
Mesaros C., Arroyo A.D., Blair I.A, & Snyder N.W. (2017). Coenzyme A thioester formation of 11- and 15-oxo-eicosatetraenoic acid. Prostaglandins & other lipid mediators, 130, 1-7.
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2

Culturing Leukemia Cell Lines and Murine Progenitors

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We cultured MV4-11 and Kasumi-1 cells in RPMI-1640 (Invitrogen, Carlsbad, USA) containing 10% heat-inactivated FBS (Gibco, Grand Island, USA), 1% penicillin-streptomycin (Sigma-Aldrich, St Louis, USA) and 1% HEPES (Sigma-Aldrich). Murine progenitor cells were cultured in RPMI 1640 containing 10 ng/ml interleukin 3 (IL-3) (Peprotech, Rocky Hill, USA), 10 ng/ml IL-6 (Peprotech), 100 ng/ml stem cell factor (SCF) (Peprotech), 55 nM 2-mercaptoethanol (BME) (Sigma-Aldrich), 1% HEPES, 10% FBS, and 1% PS. The AML patients' samples were acquired from the First Affiliated Hospital of Zhejiang University and the University of Chicago Hospital with informed consent. The study was approved by the institutional review board of two hospital's ethics committee.
Hu C., Yu M., Li C., Wang Y., Li X., Ulrich B., Su R., Dong L., Weng H., Huang H., Jiang X., Chen J, & Jin J. (2020). miR-550-1 functions as a tumor suppressor in acute myeloid leukemia via the hippo signaling pathway. International Journal of Biological Sciences, 16(15), 2853-2867.
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3

Culture of Cells in Nutrient-Deprived Media

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Complete media consisted of RPMI1640 (Mediatech) supplemented with 10% FBS (Sigma), 2mM L-glutamine (Sigma; final concentration 4mM), 0.05mM 2-mercaptoethanol (BME, Sigma), and antibiotics (penicillin/streptomycin). Glutamine-free media was prepared with glutamine-free RPMI1640 (Sigma). Glucose-free media was prepared with a combination of PBS and glucose-free RPMI1640 (Gibco) (15% PBS, 85% RPMI) and supplemented with 10% dialyzed FBS containing <50ug/ml glucose by enzymatic assay [Glucose (HK) assay, Sigma], 0.05mM BME, 2mM L-glutamine (final concentration 4mM), and antibiotics. For culture of cells in glucose or glutamine free media, cells were washed in an excess of PBS and then brought up in the appropriate media prior to culture. Inhibitors were obtained from Sigma including oligomycin A, antimycin A, etomoxir, and 2DG, and added to cultures at the indicated concentrations. Galactose, Galactose oxidase, and H202 were obtained from Sigma and added at the indicated concentrations.
Keppel M.P., Topcagic N., Mah A.Y., Vogel T.P, & Cooper M.A. (2015). Activation-specific metabolic requirements for NK cell IFN-γ production. Journal of immunology (Baltimore, Md. : 1950), 194(4), 1954-1962.
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4

Feeder-free mESC Culture Conditions

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All experiments were done in 129P2/OlaHsd mouse embryonic stem cells (mESC), which were cultured according to previously published protocols [25 ]. mESCs were maintained on gelatin-coated plates feeder-free in mESC media composed of Knockout DMEM (Life Technologies) supplemented with 15% defined foetal bovine serum (FBS) (HyClone), 0.1 mM nonessential amino acids (NEAA) (Life Technologies), Glutamax (GM) (Life Technologies), 0.55 mM 2 -mercaptoethanol (b -ME) (Sigma), 1X E SGRO LIF (Millipore), 5 nM GSK-3 inhibitor XV and 500 nM UO126. Cells were regularly tested for mycoplasma.
Szczesnik T., Ho J.W, & Sherwood R. (2019). Dam mutants provide improved sensitivity and spatial resolution for profiling transcription factor binding. Epigenetics & Chromatin, 12, 36.
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5

Endothelial Cell Culture Protocol

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2-Mercaptoethanol (BME), CaCl2, MgCl2, buprenorphine, sodium bicarbonate, ascorbic acid, heparin sodium salt, and endothelial cell growth factor supplement were from Sigma-Aldrich (St. Louis, MO, USA). Ficoll-Paque PLUS was from GE Healthcare (Uppsala, Sweden). Penicillin/Streptomycin (P/S), PBS, RPMI, and human serum-type AB (HS) were from Corning (Corning, NY, USA). CCL2 was from PreproTech (Rocky Hill, NJ, USA) and R&D Systems (Minneapolis, MN, USA). Macrophage colony stimulating factor (MCSF-1) was from PreproTech, digitonin from Invitrogen (Carlsbad, CA, USA), EDTA from Promega (Madison, WI, USA), and Bradford protein assay was from Bio-Rad (Hercules, CA, USA). Protein A/G and protein-A agarose beads were from Santa Cruz (Dallas, TX, USA). HEPES was from Technova (Hollister, CA, USA), bovine serum albumin from Life Sciences US biological (Salem, MA, USA), paraformaldehyde (PFA) from Electron Microscopy Sciences (Hatfield, PA, USA) and nonfat dry milk was from LabScientific (Highlands, NJ, USA). Fetal bovine serum (FBS), M199, HBSS, TRIS-HCL, NaCl, calcein-AM, glycerol, Medium 199 (M199), L-glutamine, and new born calf serum were from Thermo Fischer Scientific (Waltham, MA, USA). Bovine brain extract was from Clonetics/Lonza (Walkersville, MD, USA).
Jaureguiberry-Bravo M., Lopez L, & Berman J.W. (2018). Buprenorphine decreases CCL2-mediated migration of CD14+CD16+ monocytes. Journal of leukocyte biology, 104(6), 1049-1059.
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6

Borrelia Species Protein Extraction

Cited 1 time
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Low-passaged (≤10 laboratory passages) cultures of B. burgdorferi B31 A3 (63 (link)), B. turicatae 91E135 (64 (link)), B. parkeri SLO, and B. hermsii HCT-4 (29 (link)) were grown at 35 to 37°C to densities of  >1 × 107 cells mL−1 in 40 mL modified Barbour-Stonner-Kelly (mBSK) medium with 12% rabbit serum (65 (link), 66 ). Cultures were centrifuged at 11,000 × g for 20 min at 10°C. Cells were washed twice with phosphate-buffered saline (PBS) plus 5mM MgCl2 and centrifuged following each wash. Cells were resuspended in a 1:1 solution of PBS plus 5 mM MgCl2:2× Laemmli sample buffer (SB) (Bio-Rad, Hercules, CA, USA) with 2-mercaptoethanol (BME) (Sigma, St. Louis, MO, USA) at a density of 2 × 106 cells μL−1.
Curtis M.W., Krishnavajhala A., Kneubehl A.R., Embers M.E., Gettings J.R., Yabsley M.J, & Lopez J.E. (2022). Characterization of Immunological Responses to Borrelia Immunogenic Protein A (BipA), a Species-Specific Antigen for North American Tick-Borne Relapsing Fever. Microbiology Spectrum, 10(3), e01722-21.
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7

Isolation and Stimulation of Murine CD19+ B-cells

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Splenocytes from IL-10-GFP reporter male and female mice were collected as described in (Bodhankar et al. 2014a ). Splenic CD19+ B-cells were purified using paramagnetic bead-conjugated antibodies (Abs) from the CD19 cell isolation kit and subsequently separated by AutoMACS (Miltenyi Biotec, Auburn, CA). The negative fraction of the cells thus separated were CD19+ B-cells with a purity of ≥ 95%. CD19+ B-cells were suspended in RPMI 1640 medium (Mediatech, Manassas, VA, USA) supplemented with 2% Fetal Bovine Serum (FBS), 57.2μM 2-mercaptoethanol (BME, Sigma-Aldrich), 1% L-glutamate (Life Technologies), 1% sodium pyruvate (Life Technologies), and cultured in the presence of 1 μg/mL lipopolysaccharide (LPS, E. coli strain K12). After 48 h of culture, B-cells were harvested from culture plates, washed free of LPS and counted in 0.2% trypan blue (Life Technologies) in a cell counter (Nexcelom Bioscience, Lawrence, MA, USA), washed, and resuspended in DMEM culture medium without GM-CSF for co-culture with microglia.
Bodhankar S., Lapato A., Chen Y., Vandenbark A.A., Saugstad J.A, & Offner H. (2015). Role for Microglia in Sex Differences after ischemic stroke: Importance of M2. Metabolic brain disease, 30(6), 1515-1529.
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8

Borrelia Species Protein Extraction

Cited 1 time
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Low-passaged (≤10 laboratory passages) cultures of B. burgdorferi B31 A3 (63 (link)), B. turicatae 91E135 (64 (link)), B. parkeri SLO, and B. hermsii HCT-4 (29 (link)) were grown at 35 to 37°C to densities of  >1 × 107 cells mL−1 in 40 mL modified Barbour-Stonner-Kelly (mBSK) medium with 12% rabbit serum (65 (link), 66 ). Cultures were centrifuged at 11,000 × g for 20 min at 10°C. Cells were washed twice with phosphate-buffered saline (PBS) plus 5mM MgCl2 and centrifuged following each wash. Cells were resuspended in a 1:1 solution of PBS plus 5 mM MgCl2:2× Laemmli sample buffer (SB) (Bio-Rad, Hercules, CA, USA) with 2-mercaptoethanol (BME) (Sigma, St. Louis, MO, USA) at a density of 2 × 106 cells μL−1.
Curtis M.W., Krishnavajhala A., Kneubehl A.R., Embers M.E., Gettings J.R., Yabsley M.J, & Lopez J.E. (2022). Characterization of Immunological Responses to Borrelia Immunogenic Protein A (BipA), a Species-Specific Antigen for North American Tick-Borne Relapsing Fever. Microbiology Spectrum, 10(3), e01722-21.
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9

Detecting m1A and m6A Modifications

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Reagents DMS, 2-mercaptoethanol (b-ME), echinomycin, m 3 A and m 7 G nucleobases were purchased from Sigma-Aldrich. m 1 A monoclonal antibody (mouse) was purchased from MBL. The m 1 A nucleobase was purchased from Acros Organics. Horseradish peroxidase (HRP)-conjugated secondary antibody (anti-mouse IgG) was purchased from ThermoFisher. The AlkB enzyme was a gift from Dr. Patrick O'Brien (University of Michigan). Unmodified DNA oligonucleotides were purchased from IDT with HPLC purification for primer extension assays and using standard desalting for other experiments. 5'-IR700-labeled DNA oligos were purchased from IDT with HPLC purification. DNA oligonucleotides containing m 1 A or m 6 A were purchased from Yale Keck Biotechnology.
Xu Y., Manghrani A., Liu B., Shi H., Pham U., Liu A., & Al‐Hashimi H.M. (2020). Hoogsteen base pairs increase the susceptibility of double-stranded DNA to cytotoxic damage.
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