RAW 264.7 cells (5–105 cells per well) were incubated for 24 hours in the absence and presence of 0.5 µM MBP C8 or MBP C1 isomers. In other series of experiments, RAW 264.7 cells were treated with cytokines to induce either M1 or M2 cell state with or without 0.5 µM MBP charge isomers. For M1 polarization, cells were treated for 24 hours with and without 0.5 µM MBP charge isomers in the presence of 20 ng/mL of interferon-gamma (IFN-γ) (ab123747; Abcam, Cambridge, UK) and 100 ng/mL of lipopolysaccharide (LPS) (L2880-100MG; Sigma-Aldrich Co.); for M2 polarization, in the presence of 20 ng/mL of IL-4 (ab191628; Abcam) and 10 ng/mL of IL-10 (BMS347; eBioscience, Vienna, Austria). Cell viability was assessed by staining the cells with Trypan blue (#1450021; Bio-Rad, Hercules, CA, USA) using an automated Cell Counter TC 20™ (Bio-Rad). The cells were then harvested using Cell Scraper (C6106-100EA; Greiner Bio One, Frickenhausen Germany), and the harvested cells were maintained in growth medium and used for further analysis.
L2880 100mg
Manufactured by Merck Group
Sourced in United States
L2880-100MG is a lab equipment product offered by Merck Group. It is a chemical compound used in various research and scientific applications. The core function of this product is to serve as a laboratory reagent, without further interpretation or extrapolation on its intended use.
Automatically generated - may contain errors
Lab products found in correlation
Cell scraper, by Greiner (1 mentions)
Trypan blue, by Bio-Rad (1 mentions)
Cell counter tc 20, by Bio-Rad (1 mentions)
Dm2500 optical microscope, by Leica (1 mentions)
7100 automatic biochemical analyzer, by Hitachi (1 mentions)
Chemidoc xrs gel imaging system, by Bio-Rad (1 mentions)
Electrophoresis unit, by Bio-Rad (1 mentions)
Rm2235 paraffin slicer, by Leica (1 mentions)
Lightcycler 480, by Roche (1 mentions)
Sybr green 1 master, by Roche (1 mentions)
Il 10 elisa kit, by BioLegend (1 mentions)
5 protocols using l2880 100mg
1
Modulation of Macrophage Polarization by MBP Isomers
2
Endotoxin-Induced Organ Dysfunction Study
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LPS (Lipopolysaccharides from Escherichia coli O555: B5, L2880-100MG) was purchased from Sigma (USA) (Batch number: 017M4112V). Dexamethasone (DXM) was purchased from Anhui Golden Sun Biochemical Pharmaceutical Co., Ltd. (Anhui, China) (batch number: 15032521). 7100 Automatic biochemical analyzer (Hitachi, Japan), Bio-Rad electrophoresis unit (USA), and Bio-Rad ChemiDocXRS + Gel Imaging System (USA) were used in this study. LEICA RM2235 paraffin slicer (Germany) was used to generate sections for microscopy. LEICA DM2500 Optical Microscope (Germany) was used to measure neutrophil invasion. Urea nitrogen (BUN) (R1 TG836, R2 TG837), total protein (TP) (TH619), albumin (ALB) (TF126), aspartate aminotransferase (AST) (R1 AR792, R2 TG862), alanine aminotransferase (ALT) (R1 AR794, R2 TH622), lactate dehydrogenase (LDH) (R1 AR796, R2AP318) and alkaline phosphatase (ALP) (R1 TF168, R2 AR800) were all purchased from Japan Pure Pharmaceutical Industry Co., Ltd. in Shanghai, China. Creatinine (Cre) (710241 H) and creatine kinase (CK) (708021 G) were purchased from Beijing Leadman Biochemical Co., Ltd. (Beijing, China).
3
LPS-induced Endotoxemia in C57BL/6 Mice
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LPS (Sigma, L-2880-100 MG, Burlington, MA, USA) was dissolved in phosphate buffered saline (PBS) to achieve a final concentration of 5 mg/mL and subsequently stored at −20 °C for future use. Male C57BL/6 mice, aged 8 to 12 weeks, were procured from the Laboratory Animal Center at Southern Medical University. TLR4−/−, MyD88−/−, and TRIF−/− mice were generously donated by Prof. T.R. Brilliar from the Department of Surgery at the University of Pittsburgh, Pittsburgh, PA, USA.
The mice were maintained in a specific-pathogen-free (SPF) environment, with regulated temperature and humidity levels. During the feeding process, these mice were exposed to a standard dietary regimen, unrestricted access to potable water, and alternating 12-h periods of light and darkness, while maintaining an air exchange rate of 12–15 times per hour. Prior to the study, the mice were acclimatized for a minimum of one week. To establish LPS-associated endotoxemic animal models, the mice were subjected to anesthesia through inhalation of isoflurane and intraperitoneally injected with a single lethal dose of 20 mg/kg of LPS, as per the relevant literature [64 (link),65 (link)]. The animal experiments performed in this study had received approval from the Animal Care and Use Committee of Southern Medical University, Guangzhou, China (L2018235, 17 December 2018).
The mice were maintained in a specific-pathogen-free (SPF) environment, with regulated temperature and humidity levels. During the feeding process, these mice were exposed to a standard dietary regimen, unrestricted access to potable water, and alternating 12-h periods of light and darkness, while maintaining an air exchange rate of 12–15 times per hour. Prior to the study, the mice were acclimatized for a minimum of one week. To establish LPS-associated endotoxemic animal models, the mice were subjected to anesthesia through inhalation of isoflurane and intraperitoneally injected with a single lethal dose of 20 mg/kg of LPS, as per the relevant literature [64 (link),65 (link)]. The animal experiments performed in this study had received approval from the Animal Care and Use Committee of Southern Medical University, Guangzhou, China (L2018235, 17 December 2018).
4
LPS-Induced Immune Cell Dynamics
5
Analyzing Cytokine Expression in LPS-Treated Mice
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Total mRNAs were reverse-transcribed to cDNAs and gene expression levels were determined using a Light Cycler 480 and SYBR Green I Master (04707516001; Roche Diagnostics). The values were normalized by the expression of β-actin. Specific primers used for real-time PCR are as follows: Il10 forward: 5′-GGTTGCCAAGCCTTATCGGA-3′, Il10 reverse: 5′-ACCTGCTCCACTGCCTTGCT-3′, Tbx21 forward: 5′-AATCGACAACAACCCCTTTG-3′, Tbx21 reverse: 5′-AACTGTGTTCCCGAGGTGTC-3′, Gata2 forward: 5′-CAAGAAAGGGGCTGAATGTTTCG-3′, Gata2 reverse: 5′-GTGTCCCACAGGTGCCATG-3′, Actb forward: 5′-TGTTACCAACTGGGACGACA-3′, Actb reverse: 5′-CTGGGTCATCTTTTCACGGT-3′. To evaluate the secretion of IL-10 from BM B cells, B cell populations or total BM cells were obtained from WT or Cd19-Cre Il10fl/fl mice before or 24 or 48 h after LPS treatment (L2880-100MG, 5 mg/kg; Sigma-Aldrich) and were cultured in RPMI-1640 media containing 10% FBS in the presence of LPS (100 ng/ml) for 24 h. IL-10 concentrations in the supernatants were determined using an IL-10 ELISA kit (Biolegend). Levels of TNF-α, IL-6, IL-1β, G-CSF, and GM-CSF in the serum 2 h after LPS injection (5 mg/kg) were measured using ELISA kits purchased from R&D Systems.
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