Transcriptional activation of RAR‐reporter constructs by tRA was found to be maximal between 0.1 and 10 μM (Allenby et al. 1993 ), and experiments with glial cultures showed strong effects with 0.01–1 μM (van Neerven, Regen, et al., 2010 ). To test the molecular regulation of phagocytosis we therefore tested 10 nM, 0.1 μM, and 0.5 μM of the pan‐RAR agonist all‐tRA (R2625; Sigma); 0.1 and 0.5 μM pan‐RXR agonist bexarotene (153559‐49‐0; LC Laboratories); 1 μM pan‐RXR antagonist UVI3003 (3303; Tocris); 1 μM RARβ agonist‐RARα/γ antagonist BMS189453 (SML1149; identical to BMS453; Sigma); 1 and 10 μM pan LXR agonist T0901317 (Cay71810‐10; Cayman); 9 and 18 μM PPARα agonist fenofibrate (Cay10005368‐1; Cayman); 50 nM and 0.1 μM PPARγ antagonist rosiglitazone (LKT‐R5773.100; LKT Laboratories); 0.2 and 0.5 μM PPARβ/δ agonist GW501516 (Cay10004272‐1; Cayman); 1 and 2 μM FXR agonist GW4064 (Cay10006611‐5; Cayman); 0.5 and 1 μM TGM2 inhibitor cystamine dihydrochloride (B22873.14; Alfa Aesar); 15, 25, 50, and 100 μM TGM2 inhibitor ERW1041E (5095220001; Merck); and 1, 5, and 25 μM blocker of scavenger receptor CD36 sulfo‐N‐succinimidyl oleate (SSO; SML2148; Merck).
Sml2148
Manufactured by Merck Group
The SML2148 is a laboratory equipment product manufactured by Merck Group. It is designed to perform specific functions in a laboratory setting. The core function of the SML2148 is to provide precise and reliable measurements, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Gw4064, by Cayman Chemical (2 mentions)
Cystamine dihydrochloride, by Thermo Fisher Scientific (1 mentions)
Fenofibrate, by Cayman Chemical (1 mentions)
Uvi3003, by Bio-Techne (1 mentions)
Bms189453, by Merck Group (1 mentions)
Bms453, by Merck Group (1 mentions)
Gw501516, by Cayman Chemical (1 mentions)
T0901317, by Cayman Chemical (1 mentions)
R2625, by Merck Group (1 mentions)
Bexarotene, by LC Laboratories (1 mentions)
Rosiglitazone, by LKT Laboratories (1 mentions)
Mouse ifn γ, by Thermo Fisher Scientific (1 mentions)
Escherichia coli lps, by Merck Group (1 mentions)
Bay11 7082, by Merck Group (1 mentions)
H89 dihydrochloride, by Merck Group (1 mentions)
Ab120301, by Abcam (1 mentions)
Etomoxir, by Merck Group (1 mentions)
Palmitate, by Merck Group (1 mentions)
4 protocols using sml2148
1
Molecular Regulation of Phagocytosis
2
Inflammatory Pathways and Bile Acid Modulation
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Inflammatory pathways were induced with Escherichia coli LPS (isotype O26:B6; Sigma‐Aldrich) at concentrations of 0.1, 1, 10 and 100 ng/ml and with mouse IFNγ (Peprotech) 10 and 20 ng/ml. TUDCA sodium salt was purchased from Calbiochem (580549), sodium TLCA from Sigma‐Aldrich (T7515) and HDCA from Alfa Aesar (B20506). In preliminary experiments with Raw264.7 cells, using nitrite release as the inflammatory read out, we tested concentrations of 20–320 µM bile acids, dissolved in cell culture medium. Based on these results we chose 40 µM TLCA, 200 µM TUDCA and 160 µM HDCA in phagocytosis and gene expression assays. The activation of NFκB was inhibited with 2 µM Bay11‐7082 (Sigma‐Aldrich B5556). We blocked CD36 with 25 μM sulfo‐N‐succinimidyl oleate (SSO; Merck SML2148) and PKA with 2 µM H89 dihydrochloride (Sigma‐Aldrich B1427). All‐trans retinoic acid (RA; Sigma‐Aldrich R2625) was used at 0.1 µM and GW4064 (Cayman 10006611‐5) at 2 µM (Wu et al., 2021 (link)).
3
Astrocyte Model for Amyloid-Beta Pathogenesis
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Normal human astrocytes (NHA, Lonza) were routinely grown in ABM Basal Medium (CC-3187) supplemented with AGM SingleQuotsTM Supplements (CC-4123) in a cell culture incubator (5% CO2, 37 °C). For subsequent treatments and sampling, 5000 cells/cm2 were seeded for 1 week in 6-well plates, with periodic medium change.
Beta-amyloid peptide (1–42) (ab 120301, Abcam) and the corresponding inactive control (ab120481) were reconstituted to a 200 µM stock in NaOH 10 mM/HEPES 25 mM, diluted to working concentration (0.2 µM), and incubated in complete cell medium up to 48 h in a cell culture incubator (5% CO2, 37 °C), for fibril formation.
Sulfo-N-succinimidyl Oleate sodium (SSO) (Sigma, SML2148) was reconstituted in DMSO to 5.2 mM stock concentration and diluted to working concentration (20 µM) with complete cell medium. Control cells were incubated with vehicle (complete cell medium supplemented with the respective concentration of DMSO 1/200). Cells were pre-treated with a working concentration of SSO for 10 min, then the medium was replaced with normal cell medium, with or without FAs (PA and OA).
PA (Sigma P0500) and OA (Sigma O1008) were reconstituted in 70% ethanol to a 40 mM stock concentration, which was diluted to working concentrations (OA 40 µM and PA 20 µM) with complete cell medium. Corresponding controls were incubated with vehicle (medium supplemented with 0.07% ethanol).
Beta-amyloid peptide (1–42) (ab 120301, Abcam) and the corresponding inactive control (ab120481) were reconstituted to a 200 µM stock in NaOH 10 mM/HEPES 25 mM, diluted to working concentration (0.2 µM), and incubated in complete cell medium up to 48 h in a cell culture incubator (5% CO2, 37 °C), for fibril formation.
Sulfo-N-succinimidyl Oleate sodium (SSO) (Sigma, SML2148) was reconstituted in DMSO to 5.2 mM stock concentration and diluted to working concentration (20 µM) with complete cell medium. Control cells were incubated with vehicle (complete cell medium supplemented with the respective concentration of DMSO 1/200). Cells were pre-treated with a working concentration of SSO for 10 min, then the medium was replaced with normal cell medium, with or without FAs (PA and OA).
PA (Sigma P0500) and OA (Sigma O1008) were reconstituted in 70% ethanol to a 40 mM stock concentration, which was diluted to working concentrations (OA 40 µM and PA 20 µM) with complete cell medium. Corresponding controls were incubated with vehicle (medium supplemented with 0.07% ethanol).
4
Fatty Acid Uptake Protocol
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Here, 100 μM, 250 μM, and 500 μM palmitate (Sigma-Aldrich, catalog no. P9767) and oleate (Fisher Scientific, catalog no. 75096) conjugated to fatty acid-free BSA were used for feeding cells in incomplete LG media (63 ). Fat-free BSA was used as a control. Etomoxir treatment (Sigma-Aldrich, catalog no. E1905) was given at a concentration of 60 μM for 16 h to inhibit CPT1 (68 (link)). SSO (Sigma-Aldrich, catalog no. SML2148) was used to inhibit CD36 at a concentration of 50 μM for 12 h. Fatty acid uptake was detected using LC-MS/MS as subsequently described.
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