The exopolysaccharides (EPS) (2 MDa) were obtained by the blue-green cynobacteria Neocyanospira capsulata, a strain of which was present in the culture collection of the Department of Agriculture, Food, Environment and Forestry (Microbiology Section) of the University of Florence, following the methodology described by Vincenzini et al. (1990 (link)). Egg phosphatidylcholine (EPC) was purchased from Avanti Polar Lipids Inc., Alabaster, AL. Capsaicin (CAP), with Molecular Weight (MW) of 305.41 g/mol, was purchased by Merck (Darmstadt, Germany) as well as all the solvents used. Hydroxytyrosol (HT), with MW of 154.16 g/mol, was extracted from the free-oil olive pulp at the PHYTOLAB, University of Florence, following the procedure described by Romani et al. (2017 (link)).
Capsaicin cap
Manufactured by Merck Group
Sourced in United States
Capsaicin (CAP) is a naturally occurring chemical compound found in certain types of peppers. It is a colorless, crystalline substance that is the primary active component responsible for the pungent sensation experienced when consuming chili peppers. CAP is commonly used in research and laboratory settings to study its physiological and biochemical properties.
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Lab products found in correlation
Egg phosphatidylcholine epc, by Avanti Polar Lipids (1 mentions)
Diethylnitrosamine den, by Merck Group (1 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
P acc ser79, by Cell Signaling Technology (1 mentions)
Anti caspase 3 antibody, by Santa Cruz Biotechnology (1 mentions)
Sorafenib, by Merck Group (1 mentions)
P ampkα1 thr172, by Cell Signaling Technology (1 mentions)
Anti caspase 9, by Cell Signaling Technology (1 mentions)
Peroxidase labeled secondary anti mouse igg, by Merck Group (1 mentions)
Anti afp, by Cell Signaling Technology (1 mentions)
Anti p akt ser473, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Complete freund s adjuvant cfa, by Merck Group (1 mentions)
Serotonin hydrochloride 5 ht, by Merck Group (1 mentions)
β estradiol e2, by Merck Group (1 mentions)
M100907, by Merck Group (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Amphotericin b, by Thermo Fisher Scientific (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Sto 609, by Merck Group (1 mentions)
7 protocols using capsaicin cap
1
Extraction and Characterization of Cyanobacterial EPS
2
Diethylnitrosamine and Capsaicin Protocol
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Diethylnitrosamine (DEN, Sigma-Aldrich, Missouri, USA) was dissolved in saline (0.9% NaCl). Capsaicin (CAP, Merck Millipore, Missouri, USA) was first dissolved in ethanol to a stock solution and this was further dissolved in saline to form a final solution containing 0.1 or 0.05 mg CAP in 1 ml 1% ethanol solution (dosage 1.5 mg/kg body weight [BW] or 0.75 mg/kg BW). The doses of capsaicin were set up according a previous study (Díaz Barriga et al. 1995) .
3
Apoptosis and Proliferation Signaling
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Capsaicin (CAP) and Sorafenib were purchased to Sigma (St. Louis, MO, USA). Primary antibodies anti-caspase-9, anti-PARP, anti-AFP, anti-pAkt-ser473, p-mTOR-ser2448, p-AMPKα1-thr172, p-ACC-ser79 and the antibodies against the corresponding total forms were obtained from Cell Signaling Technology (Danvers, MA, USA). The anti-caspase-3 antibody was from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). (Peroxidase labeled secondary anti-mouse IgG was from Sigma (St. Louis, MO, USA) and anti-rabbit IgG was from Calbiochem (San Diego, CA, USA).
4
Preparation of Serotonin, Capsaicin, and Associated Compounds
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Serotonin hydrochloride (5HT; Sigma–Aldrich, St. Louis, MO) was dissolved in double-distilled water and diluted in 0.9% sterile saline or Hank’s balanced salt solution (HBSS) buffer immediately prior to each use. Capsaicin (CAP; Sigma–Aldrich, St. Louis, MO) was dissolved in 100% ethanol in a fume hood and aliquots were stored at −20 °C as 100 mM stocks. Capsaicin was freshly diluted in 0.9% saline or HBSS buffer prior to each use. The 5HT2A antagonist, M100907 (Sigma–Aldrich), was dissolved in 20% dimethyl sulfoxide (DMSO) in 0.9% sterile saline, stored as a 2 mM stock solution at 4 °C, and then serial diluted the day of use to a final working solution of 2 nM, 10 nM, or 30 nM M100907 (6% DMSO in 0.9% sterile saline). β-Estradiol (E2; Sigma–Aldrich, St. Louis, MO) was dissolved in 100% ethanol to create a 10 mM stock solution that was further diluted in HBSS buffer for a working solution of 50 nM. Immediately prior to use, complete Freund’s adjuvant (CFA; Sigma–Aldrich) was dissolved 1:1 in 0.9% sterile saline.
5
Fecal Microbiome Cultivation Protocol
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The starting homogenate of each individual human fecal sample (Microbiome Health Research Institute, Boston, MA, USA) was harvested from a single American, randomly selected as described previously [31 (link),32 (link)], for a total of 2 individual human fecal samples. The pancreatic juice (PJ) and defined medium (DM) were prepared as described previously [32 (link)]. Capsaicin (CAP) was purchased from Sigma Aldrich (St. Louis, MO, USA).
6
Capsaicin and Capsazepine Effects on LPS-induced Neuroinflammation
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Capsaicin (CAP) was obtained from Sigma, and capsazepine (CZP) from Tocris (Ellisville, USA). Animals were treated with CAP (0.001–2.5 mg/kg, intraperitoneal (i.p.) injection) 30 min before LPS intranigral injection, and CZP (1 mg/kg, i.p.) was administered 30 min before CAP. Control animals received an intranigral injection of vehicle (PBS). CAP and CZP were dissolved in a vehicle consisting of an 8:1:1 ratio of PBS:TWEEN 80:absolute ethanol26 (link). There was no difference in body weight or growth rate between any of the groups.
7
Prostate cancer cell culture protocol
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Capsaicin (CAP) and STO-609 were purchased to Sigma-Aldrich (St. Louis, MO, USA). psPAX2 vector was a gift from Didier Trono (Addgene plasmid #12260; http://n2t.net/addgene:12260; Cell culture. PC3, DU-145 and LNCaP human prostate cancer cell lines were obtained from American Type Culture Collection, (ATCC CRL-1435, ATCC HTB-81 and ATCC CRL-1740 respectively) (Rockville, MD, USA). Cells were routinely grown in RPMI 1640 medium supplemented with 100 IU/ml penicillin G sodium, 100 mg/ml streptomycin sulfate, 0.25 mg/ml amphotericin B (Invitrogen, Paisley, UK) and 10% fetal bovine serum. All cell lines were incubated at 37°C in 5% CO 2 and routinely tested for Mycoplasma infection. For treatment experiments, cells were plated and grown 24h, the medium was then replaced with serum-free RPMI 1640 and then incubated with different treatments for the indicated times. Cells were used at passages 4-20.
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