Mice were fed the HFD without guar gum for 6 weeks. Cecal SCFA infusions were performed as described previously [22 (link)]. Briefly, mice were equipped with a permanent cecum catheter and allowed a recovery period of at least 5 days. On the day of the experiment, the mice were individually housed and fasted from 6:00 to 10:00 a.m. All infusion experiments were performed in conscious, unrestrained mice. Four different groups of each 8 mice received a 140 mM phosphate-buffered saline (140 mM NaCl, 10 mM sodium phosphate at pH 5.8), a 140 mM sodium acetate (S2889; Sigma), a 140 mM sodium propionate (P1880; Sigma) or a 140 mM sodium butyrate (303410; Sigma) solution infused via the cecum catheter at a rate of 0.2 ml/h for 6h. The infusion rate of SCFA was based on the recommended intake of dietary fiber for humans of 38 g/day/human, which results in approximately 380 mmol SCFAs/day/human [23 ,24 (link)]. When calculated for mice, this corresponds to 170 μmol SCFAs/day/mouse. By infusing 140 mM SCFA directly into the cecum at a rate of 0.2 ml/h for 6h, a total amount of 168 μmol SCFA was given per mouse. After 6h of infusion, animals were terminated by cardiac puncture under isoflurane anesthesia. Cecum was removed quickly, freeze-clamped, and stored at -80°C. Blood was centrifuged (4000 x g for 10 min at 4°C) and plasma was stored at -80°C.
P1880
Manufactured by Merck Group
Sourced in United States
The P1880 is a laboratory equipment product from Merck Group. It is designed for general laboratory use, but a detailed description of its core function is not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
S2889, by Merck Group (5 mentions)
B5887, by Merck Group (3 mentions)
S9888, by Merck Group (1 mentions)
Goat anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions)
L glutamine, by Merck Group (1 mentions)
Gentamicin, by Merck Group (1 mentions)
Flat bottomed 96 well plate, by Thermo Fisher Scientific (1 mentions)
Soluble anti cd28 mab, by BD (1 mentions)
S5636, by Merck Group (1 mentions)
Celldiscoverer 7, by Zeiss (1 mentions)
P0500, by Merck Group (1 mentions)
V900933, by Merck Group (1 mentions)
6 aminocaproic acid, by Merck Group (1 mentions)
D4902, by Merck Group (1 mentions)
A2504, by Merck Group (1 mentions)
9 protocols using p1880
1
SCFA Infusion and Metabolic Phenotyping in Mice
2
SCFA Mixture Supplementation After Surgery
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After undergoing surgery, the rats were given a mixture of SCFAs with 67.5 mM sodium acetate (S2889, Sigma), 25.9 mM sodium propionate (P1880, Sigma), and 40 mM sodium butyrate (303410, Sigma), or a salt-matched control solution (133.4 mM sodium chloride) (S9888, Sigma) in drinking water ad libitum continuously except for during the SPT [75 (link)].
3
Sodium Propionate Impact on Mitochondria
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4
Short-chain Fatty Acid Mixture Administration
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A mixture of SCFA was administered according to previous reports [48 (link), 82 (link), 83 (link)]. SCFA mixture was composed of 67.5 mM of acetate (S2889, Sigma-Aldrich), 25.9 mM of propionate (P1880, Sigma-Aldrich), and 40 mM of butyrate (303410, Sigma-Aldrich). The mixture was diluted in the Abx solution. Solutions with no SCFA mixture were sodium matched. All solutions were pH controlled (7.4–7.6) and changed weekly.
5
Optimizing PBMC Proliferation Assays
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Total PBMC or LP cell populations (2 × 105 cell/well) were cultured for 3 or 2 days, respectively, at 37 °C and 5% CO2, in Iscove’s medium supplemented with 10% human FBS, 100 µg/mL gentamicin (Sigma-Aldrich) and 3 µg/mL L-glutamine (Sigma-Aldrich), without or with different concentrations of sodium acetate (S2889), sodium proprionate (P1880) or sodium butyrate (B5887, all from Sigma Aldrich). The cell cultures were stimulated polyclonally in flat-bottomed 96-well plates (Nunc, Roskilde, Denmark). Plates were precoated with 3 µg/mL goat anti-mouse IgG (Jackson Immunoresearch labs, West Grove, PA, USA), and 12.5 ng/mL mouse anti-CD3mAb (BD Pharmingen, San Diego, CA, USA) and 0.05 µg/mL soluble anti-CD28mAb (BD Pharmingen) were added together with the cells. T cell proliferation was measured by dilution of 5, 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) dye. CFSE (Invitrogen) was added to 1 × 106 cells/mL PBMCs in a final concentration of 0.2 µM and incubated at 37 °C for 10 min prior to polyclonal stimulation. Cells were immediately washed in Iscove’s medium supplemented with 10% human FBS, 100 µg/mL gentamicin and 3 µg/mL L-glutamine after the CFSE incubation period. Proliferating cells were defined as cells that had undergone at least one cell division using non-stimulated cells as control.
6
Preparing Acetate, Propionate, Butyrate, and Methadone Solutions
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Acetate (Sigma Aldrich, S5636), propionate (Sigma Aldrich, P1880), butyrate (Sigma Aldrich, B5887), and methadone (Sigma Aldrich, M0267) were reconstituted in ultrapure distilled water, sterilized through a 0.2 μm filtered syringe, and serially diluted in EMEM containing 10% FBS (Fisher Scientific, MT35016CV).
7
Cell Migration Assay with Paw Fibroblasts
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Two-well cell culture-inserts (IBIDI, 80209, Munich, Germany) were disinfected for 20 min in 70% ethanol. After having fully dried, the inserts were carefully inserted into a 12-well cell culture plate. 25,000 SFs from naïve mouse paws (passage 4) were seeded into each well of the inserts in medium ±250 µM PA (Sigma-Aldrich, P1880, St. Louis, MO, USA). Inserts were carefully removed after 12 h, and 1 mL of the respective medium (±250 µM PA) was added. Cell migration was monitored for 72 h in 15 min intervals by live cell imaging with an automated microscope (Zeiss CellDiscoverer 7, Jena, Germany).
8
Preparation of Palmitic Acid Stock Solution
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The stock solution of 100 mM PA (P0500, Sigma-Aldrich) was prepared as per a previous study (Cousin et al., 2001) (link). Briefly, PA was dissolved in 0.1 M NaOH at 70°C and then complexed with 10% fatty acid-free bovine BSA (V900933, Sigma-Aldrich) at 55°C for 10 min to achieve a final concentration of 100 mM. The stock solution of 100 mM PA was diluted in RPMI-1640 basic medium containing 2% BSA to prepared working solution. An equivalent amount of BSA was administered to the control group and all the other treatments that were compared with PA. Propionate (P1880, Sigma-Aldrich) was dissolved in RPMI-1640 basic medium to achieve a final concentration of 100 mM.
9
Embryonic Development Modulation
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