Lps sm ultrapure

Manufactured by InvivoGen

Sourced in United States

LPS-SM ultrapure is a highly purified lipopolysaccharide (LPS) product from InvivoGen. It is produced using a specific purification process to ensure low levels of contaminants. The core function of LPS-SM ultrapure is to serve as a Toll-like receptor 4 (TLR4) agonist for research applications.

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Lab products found in correlation

L lactic acid, by Merck Group (1 mentions) M4655, by Merck Group (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Sodium l lactate, by Merck Group (1 mentions) Gm csf, by ProSpec (1 mentions) Raw 264, by RIKEN BioResource Center (1 mentions) Ly294002, by Cell Signaling Technology (1 mentions) Rapamycin, by Selleck Chemicals (1 mentions) Mitotracker cmxros red fluorescent dye, by Cell Signaling Technology (1 mentions) Triciribine, by Cayman Chemical (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Neurobasal medium, by Thermo Fisher Scientific (1 mentions) Poly l ornithine, by Merck Group (1 mentions) Pam3csk4, by InvivoGen (1 mentions)

Spelling variants of this product

Ultrapure LPS (113 citations) LPS-SM (3 citations)

4 protocols using lps sm ultrapure

Differentiation and Stimulation of BMDCs and BMDMs

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B16-F1 (Riken BRC, Ibaraki, Japan) cells were grown in D-MEM (High Glucose, 043-30085, FUJIFILM Wako, Osaka, Japan) containing 10% fetal bovine serum (FBS). RAW 264.7 (Riken BRC) cells were grown in MEM (Minimum Essential Medium Eagle, M4655, Sigma-Aldrich, St. Louis, MO, USA) containing 10% FBS. BMDCs were generated as described previously (Kanemaru et al., 2017 (link)). Briefly, mouse bone marrow cells were obtained from femurs and differentiated into DCs in MEM with 10% FBS containing 10 ng/ml granulocyte macrophage colony-stimulating factor (G M-CSF; Prospec, East Brunswick, NJ, USA). Medium was replaced with fresh medium containing G M-CSF every two days. On day 6, cells were collected. BMDMs were also generated as described previously (Kanemaru et al., 2017 (link)). Bone marrow cells were obtained and incubated in MEM with 10% FBS containing 3 ng/ml macrophage colony-stimulating factor (M-CSF, PeproTech, Rocky Hill, NJ, USA). Medium was changed on day 3. On day 5, macrophages were harvested. BMDCs and BMDMs were stimulated with LPS (100 ng/ml; LPS-SM ultrapure, InvivoGen, San Diego, CA, USA) and/or L-(+)-Lactic acid (Sigma-Aldrich), and/or sodium L-lactate (Sigma-Aldrich).

Kanemaru H., Mizukami Y., Kaneko A., Tagawa H., Kimura T., Kuriyama H., Sawamura S., Kajihara I., Makino K., Miyashita A., Aoi J., Makino T., Masuguchi S., Fukushima S, & Ihn H. (2021). A mechanism of cooling hot tumors: Lactate attenuates inflammation in dendritic cells. iScience, 24(9), 103067.

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Trained Immunity in Monocytes

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Following isolation, monocytes were counted and re-suspended in a Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with L-glutamine, penicillin, streptomycin and 10% autologous serum at a concentration of 1 million monocytes/ml. After a 1h resting phase, monocytes were stimulated with BCG or RPMI (negative control) in duplicate: one singlicate was used to study primary innate responses and supernatants were filtered and harvested after 24h for cytokine measurements and lactate measurements. The other singlicate was used to study trained immunity: After 24h, BCG was washed out with a calcium- and magnesium-free PBS [PBS(−)], medium was replenished, and BCG-trained or control monocytes were incubated at 37°C for 5 days with an interim medium replenishing step on Day 3 of culture. On Day 6, supernatants were collected and a second stimulus, lipopolysaccharide from Salmonella minnesota (LPS-SM Ultrapure, Invivogen; San Diego, CA) was added. After 24 h incubation, supernatants were harvested on Day 7 of culture for cytokine and lactate measurements (Figure 1).

Angelidou A., Diray-Arce J., Conti M.G., Netea M.G., Blok B.A., Liu M., Sanchez-Schmitz G., Ozonoff A., van Haren S.D, & Levy O. (2021). Human Newborn Monocytes Demonstrate Distinct BCG-Induced Primary and Trained Innate Cytokine Production and Metabolic Activation In Vitro. Frontiers in Immunology, 12, 674334.

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Modulating B and T Cell Proliferation

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Purified naive B cells labeled with CFSE, CTV, or eFluor670 cell proliferation dyes were cultured in lipopolysaccharide (LPS-SM ultrapure; Invivogen; 10–25 μg/mL) in 48 well plates at 2.5 × 10⁵ cells/mL of culture media in a 37 °C humidified environment with 5% CO₂. To activate CD8⁺ T cell in vitro, wild-type or P14⁺ CD8⁺ T cells were purified, labeled with cell proliferation dye and stimulated with immobilized anti-CD3 (5 μg/ml), soluble anti-CD28 (1 μg/ml) and recombinant IL-2 (30 IU), or by gp33 (1 μg/ml; Anaspec) peptide-loaded splenocytes, respectively. Memory CD8⁺ T cells were harvested at 120⁺ days after LCMV infection.
Small molecule inhibitors of PI3K (LY294002; Cell Signaling), pan-AKT (Triciribine; Cayman Chemical Company), mTOR (Rapamycin; Selleckchem), and FoxO1 (AS1842856; Calbiochem) were added to the cells at the time of activation. Mitochondria membrane potential was measured by incubating cells with MitoTracker CMXRos red-fluorescent dye (Cell Signaling) for 30 minutes followed by washing cells three times with ice-cold culture media.

Lin W.H., Adams W.C., Nish S.A., Chen Y.H., Yen B., Rothman N.J., Kratchmarov R., Okada T., Klein U, & Reiner S.L. (2015). Asymmetric PI3K Signaling Driving Developmental and Regenerative Cell Fate Bifurcation. Cell reports, 13(10), 2203-2218.

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Primary Hippocampal Neuron Culture and Stimulation

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Primary hippocampal neurons were prepared for culturing, as previously described. 20 Briefly, the hippocampi were dissected and dissociated from 18-day-old fetal mice. Dissociated hippocampal neurons were plated on poly-Lornithine (Sigma) coated 6-well plates with 2% B27 (Invitrogen, San Diego, CA) and 2 mmol/L L-glutamine (Nacalai) in Neurobasal medium (Invitrogen). Two days later, 10 mmol/L cytosine arabinoside (Nacalai) was added to inhibit nonneuronal growth. After 14 days, hippocampal neurons were stimulated with 1 mg/mL Pam3CSK4 (Invivogen) and 1 mg/mL LPS-SM Ultrapure (Invivogen) in the presence (50 ng/mL, 5 mg/mL) or absence of HA4 for 6 hours on day 14.

Sunabori T., Koike M., Asari A., Oonuki Y, & Uchiyama Y. (2016). Suppression of Ischemia-Induced Hippocampal Pyramidal Neuron Death by Hyaluronan Tetrasaccharide through Inhibition of Toll-Like Receptor 2 Signaling Pathway. The American journal of pathology, 186(8).

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