Neutrophils were isolated from peripheral blood using an EasySep direct human neutrophil isolation kit (Stemcell, Vancouver, BC). Cells were then counted and cultured at 107 cells/mL in RPMI supplemented with 5% fetal bovine serum and 1% penicillin/streptomycin. Neutrophil cultures were treated with GM-CSF (1–25 ng/mL, R&D Systems, Minneapolis, MN), FSTL1 (100–1000 ng/mL, R&D Systems), Interferon-γ (IFNγ) (10 ng/mL, R&D Systems), lipopolysaccharide (LPS) (1 μg/mL, Enzo Life Sciences, Farmingdale, NY), Interleukin 4 (IL-4) (20 ng/mL, R&D Systems), Interleukin 13 (IL-13) (20 ng/mL, R&D Systems), Interleukin 25 (IL-25) (10–100ng/mL, R&D Systems), Interleukin 33 (IL-33) (10–100 ng/mL, R&D Systems), thymic stromal lymphopoietin (TSLP) (10–100 ng/mL, R&D Systems) or Leukotriene C4 (LTC4) (10−6 –10−7 Cayman Chemical, Ann Arbor, MI) for 20 hours, cell culture supernatants were collected for protein analysis and cell lysates were collected to isolate RNA. Plates were coated with E-selectin, ICAM (50ng/well, Peprotech, Rocky Hill, NJ) or bovine serum albumin (BSA) (50ng/well, Sigma Aldrich) in pH 8 tris buffered saline (TBS) overnight at 4 degrees. Neutrophils were seeded into the wells and the culture supernatants were harvested at 20 hours.
Leukotriene c4 ltc4
Manufactured by Cayman Chemical
Sourced in United States
Leukotriene C4 (LTC4) is a lab product offered by Cayman Chemical. LTC4 is a lipid mediator that is involved in various biological processes. It is used for research purposes in the study of inflammatory and allergic responses.
Automatically generated - may contain errors
Lab products found in correlation
Easysep direct human neutrophil isolation kit, by STEMCELL (1 mentions)
Interferon γ ifnγ, by R&D Systems (1 mentions)
Fstl1, by R&D Systems (1 mentions)
Interleukin 4 il 4, by R&D Systems (1 mentions)
Interleukin 13 il 13, by R&D Systems (1 mentions)
E selectin, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Gm csf, by R&D Systems (1 mentions)
Lipopolysaccharide lps, by Enzo Life Sciences (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Phospho syk, by Cell Signaling Technology (1 mentions)
Cyclooxygenase 2 cox2, by Cell Signaling Technology (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Dinitrophenyl dnp ige, by Merck Group (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Dulbecco s minimum essential medium dmem, by Thermo Fisher Scientific (1 mentions)
Erk1 2, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
3 protocols using leukotriene c4 ltc4
1
Neutrophil Isolation and Stimulation
2
Inflammatory Signaling in Mast Cells
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ME was purchased from the National Institute for Food and Drug Control (Beijing, China; purity, ≥99.5%). Dulbecco's minimum essential medium (DMEM), penicillin, streptomycin, and fetal bovine serum (FBS) were purchased from GIBCO (Grand Island, NY, USA). 4-[3-(4-Iodophenyl)-2-4(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate (WST-1) was obtained from Dojindo (Kumamoto, Japan). Specific antibodies against phospho-Lyn, Lyn, phospho-Syk, Syk, phospho-ERK1/2, ERK1/2, phospho-p38, p38, phospho-JNK, JNK, cytosolic phospholipase A2 (cPLA2), phospho-cPLA2, cyclooxygenase-2 (COX-2), and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). Specific antibodies against phospho-5-lipoxygenase (5-LO) and 5-LO, and enzyme immunoassay (EIA) kits for prostaglandin E2 (PGE2), prostaglandin D2 (PGD2), leukotriene B4 (LTB4), and leukotriene C4 (LTC4) were purchased from Cayman Chemical (Ann Arbor, MI, USA). The enzyme-linked immunosorbent assay (ELISA) kits for TNF-α and IL-4 were obtained from Bangyi Technologies Inc. (Shanghai, China). Dinitrophenyl- (DNP-) IgE was obtained from Sigma-Aldrich (St Louis, MO, USA), and DNP-bovine serum albumin (BSA) was obtained from Biosearch Technologies Inc. (Novato, CA, USA). All other chemicals were of analytical grade and were purchased from Sigma-Aldrich.
3
Inhibition of Calcium Signaling Pathways
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Cyclopiazonic acid (CPA) was purchased from Calbiochem. Arachidonic acid was obtained from MP Biomedicals. Thapsigargin and SK&F 96365 (SKF) were purchased from Tocris Bioscience. Ketoprofen, ethylene glycol tetraacetic acid (EGTA) and 2-aminoethoxydiphenyl borate (2-APB) were obtained from Sigma. Leukotriene C4 (LTC4) and Nordihydroguaiaretic acid (NDGA) were purchased from Cayman chemical.
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