Stock solutions of specific PFKFB3 inhibitor 1-(4-Pyridinyl)-3-(2-quinolinyl)-2-propen-1-one (PFK15) were made up to 30 mM in dimethyl sulfoxide (DMSO, both Sigma). PFK15 was further diluted in DMEM for experimentation. For calcium imaging experiments, PFK15 stock solutions were diluted directly in HEPES-buffered physiological saline solution (HEPES-PSS: 138 mM NaCl, 4.7 mM KCl,10 mM HEPES, 5.5 mM glucose, 1.28 mM CaCl2, 0.56 mM MgCl2, pH 7.4). Calcium-free HEPES-PSS (0Ca-HEPES-PSS: 138 mM NaCl, 4.7 mM KCl,10 mM HEPES, 5.5 mM glucose, 1.28 mM CaCl2, 0.56 mM MgCl2, 1 mM EGTA pH 7.4) and high calcium HEPES-PSS (138 mM NaCl, 4.7 mM KCl,10 mM HEPES, 5.5 mM glucose, 20 mM CaCl2, 0.56 mM MgCl2, pH 7.4) were used for clearance assays. All drugs used in the metabolic inhibitor cocktail: iodoacetate 2 mM, (IAA), bromopyruvate 200 μM (BrPy), oligomycin 10 μM (OM) and carbonyl cyanide 3-chlorophenylhydrazone 4 μM (CCCP) were obtained from Sigma-Aldrich and stock solutions dissolved in milliQ water (IAA) or DMSO.
Bromopyruvate
Manufactured by Merck Group
Sourced in Italy
Bromopyruvate is a chemical compound used in research and laboratory settings. It is a white crystalline solid that serves as a substrate for various enzymes and biochemical reactions. The core function of Bromopyruvate is to facilitate and enable specific laboratory experiments and analyses, but a detailed description of its intended use is not available in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Oligomycin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Fitc conjugated goat anti mouse igg, by Merck Group (1 mentions)
Alexa fluor 555 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Lc3 rabbit polyclonal antibody, by Merck Group (1 mentions)
P p53 ser 15 rabbit polyclonal, by Cell Signaling Technology (1 mentions)
Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions)
Butyrate, by Merck Group (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Capricorn (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
4 protocols using bromopyruvate
1
PFKFB3 Inhibitor PFK15 Protocol
2
Mitochondrial Modulation and Autophagy Assay
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Bromopyruvate (3BP), lonidamine (LND), mitochondrial inhibitors, respiratory substrates, chemicals, LC3 rabbit polyclonal antibody, α-tubulin and β-tubulin mouse monoclonal antibodies, and goat anti-mouse FITC-conjugated IgG were from Sigma (Milan, Italy). Cyt c mouse monoclonal antibody, AIF goat polyclonal antibody, β-actin goat polyclonal antibody, p53 (FL-393) rabbit polyclonal antibody, p-p53 (Ser-315) rabbit polyclonal antibody, HK-II (C-14) goat polyclonal antibody, beclin 1 (E-8) mouse monoclonal antibody, goat anti-mouse HRP-conjugated IgG, goat anti-rabbit HRP-conjugated IgG, and rabbit anti-goat HRP-conjugated IgG were from Santa Cruz Biotechnology; VDAC1 rabbit polyclonal antibody was from Calbiochem. p-p53 (Ser-15) rabbit polyclonal, SQSTM1/p62 rabbit polyclonal, Akt rabbit polyclonal, and p-Akt (Ser-473) rabbit monoclonal antibodies were from Cell Signaling Technology. LC3 mouse monoclonal antibody was from NanoTools. Mito-tracker red CM-XRos, Alexa Fluor® 488 goat anti-rabbit IgG, and Alexa Fluor® 555 goat anti-mouse IgG were Molecular Probes products.
3
THP-1 and PBMC Responses to LPS, Bromopyruvate, and Butyrate
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THP-1 cells were cultured in RPMI1640 (Thermo Fisher Scientific, Waltham, MA, USA) containing 10% fetal bovine serum (Capricorn Scientific, Auf der Lette, Germany), penicillin (100 U/ml, Thermo Fisher Scientific), and streptomycin (100 µg/ml, Thermo Fisher Scientific). PBMCs were isolated using Ficoll-Paque Plus (GE Healthcare, Chicago, IL, USA). THP-1 cells (1×106/ml) were stimulated with 100 ng/ml lipopolysaccharide (LPS) (Sigma-Aldrich, St Louis, MO, USA) in the presence or absence of various concentrations of bromopyruvate (Sigma-Aldrich) or butyrate (Sigma-Aldrich). PBMCs (1×106/ml) were stimulated with 100 ng/ml LPS in the presence or absence of 80 µM of bromopyruvate or 10 mM of butyrate.
4
Synthesis of Chitosan-Based Nanoparticles
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Chitosan oligosaccharide (Molecular Weight (MW) 5kDa), oleic acid, and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC), dimethyl sulfoxide (DMSO), bromopyruvate, bovine serum albumin, folic acid, trypan blue, crystal violet, and ethiduim bromide were purchased from Sigma-Aldrich (Milan, Italy).
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