Somatostatin

Manufactured by Merck Group

Sourced in United States, Argentina

Somatostatin is a synthetic peptide hormone that functions as an inhibitor of various endocrine and exocrine secretions. It is primarily used as a laboratory reagent for research and experimental purposes.

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Lab products found in correlation

Close spelling variants of this product

Rat anti-somatostatin (13 citations) Anti-somatostatin (6 citations) Rat monoclonal anti-somatostatin antibody (4 citations) Somatostatin‐14 (4 citations) Somatostatin (SST; (3 citations) Rabbit anti-somatostatin (3 citations)

31 protocols using somatostatin

Organoid Size Regulation by Secretagogues

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Images of CLC organoids were taken using 5X magnification before and following the addition of secretin (100nM, Sigma Aldrich), somatostatin (100nM, Sigma Aldrich), octrotide (100nM, Sigma Aldrich) or embryo transfer water serving as a negative control, at 0.5 - 2 minute intervals until organoid size stabilized. To explore the impact of octreotide on the effect of secretin, cells were pre-incubated for 30 minutes with octreotide. 100nM of secretin (Sigma Aldrich) was subsequently added to the medium and the experiment was carried out as described above. To assess the effect of VX809 on organoid size images were taken before and 6 hours following the addition of VX809 (30mM, Selleck) or embryo transfer water, serving as a negative control. 3 random diameters were measured for 8 random organoids pre and post treatment. Graph measurements represent percentage differences in mean organoid diameter. Error bars represent SD. Statistical significance was calculated using one-way ANOVA with Dunnett correction for multiple comparisons. The videos available as online supplementary data were made by taking images pre and post treatment at 2 minute intervals, until organoid size stabilized.

Sampaziotis F., de Brito M.C., Madrigal P., Bertero A., Saeb-Parsy K., Soares F.A., Schrumpf E., Melum E., Karlsen T.H., Bradley J.A., Gelson W.T., Davies S., Baker A., Kaser A., Alexander G.J., Hannan N.R, & Vallier L. (2015). Cholangiocytes derived from human induced pluripotent stem cells for disease modeling and drug validation. Nature biotechnology, 33(8), 845-852.

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Comprehensive Sweetener Research Protocol

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The following compounds and toxin were purchased from Sigma-Aldrich (St. Louis, MO, United States): Acesulfame K, aspartame, D-fructose, sucrose, sodium saccharin, monellin, thaumatin, rebaudioside A, lactisole, somatostatin, epidermal growth factor (EGF), SFLLR, Pertussis toxin (PTx), latrunculin A, wortmannin, GW583340, Tyrphostin AG 1478 and GF109203x. Stevioside was from Emperors Herbologists (Jacksonville, FL, United States). P-4000, SC-45647 and superaspartame were gifts from Grant Dubois (the Coca-Cola Company, Atlanta, GA). Sucralose was from Toronto Research Chemicals, Inc. (North York, Ontario, Canada). Alitame (Aclame™) was a generous gift from Danisco (Terre Haute, IN, United States). Neotame was from the NutraSweet Company (Chicago, IL, United States). Dulcin was from Maybridge Chemical Company (Cornwall, United Kingdom). S1P, U1026 and GSK269962 were from Tocris Biosciences (Minneapolis, MN). S819, S5227 (Zhang et al., 2008 (link)), S679 (Tachdjian et al., 2009 ) and S1313 (Tachdjian et al., 2010 ) were synthesized in house.

Servant N.B., Williams M.E., Brust P.F., Tang H., Wong M.S., Chen Q., Lebl-Rinnova M., Adamski-Werner S.L., Tachdjian C, & Servant G. (2022). A Dynamic Mass Redistribution Assay for the Human Sweet Taste Receptor Uncovers G-Protein Dependent Biased Ligands. Frontiers in Pharmacology, 13, 832529.

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Synthesis and Characterization of S-modified Peptides

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Methanol and glacial acetic acid were purchased from Mallinckrodt (Phillipsburg, NJ, USA). Sodium periodate, sodium persulfate, lysozyme, somatostatin, trypsin, pepsin, insulin, cysteine, methionine, S-nitrosoglutathione, and ammonium bicarbonate were purchased from Sigma-Aldrich (St. Louis, MO, USA). ARACAKA was synthesized by Pepnome Ltd. (Shenzhen, China). S-Glutathionylated ARACAKA was prepared by incubating 1 mg of ARACAKA with 1 mg of S-nitrosoglutathione in 1 mL of water at 37 °C for 1 h. S-cysteinylated ARACAKA was prepared by combining 1 mg of ARACAKA with 1 mg of cysteine in water for 1 week at room temperature. All peptide stock solutions for positive nanoelectrospray (nESI) were prepared in a 49.5/49.5/1 (vol/vol/vol) solution of Methanol/water/acetic acid at an initial concentration of ~1 mg/mL and diluted 100-fold prior to use. Solutions of sodium persulfate (aqueous) and Sodium periodate (50/50 vol/vol Methanol/water) anions were prepared at concentrations of ~1 mg/mL and diluted 10-fold prior to use. Sodium periodate was incubated with 1 mg/mL of methionine (50/50 vol/vol Methanol/water) to yield the iodate (IO₃⁻) anion.

Pilo A.L, & McLuckey S.A. (2016). Selective Gas-Phase Ion/Ion Reactions: Enabling Disulfide Mapping via Oxidation and Cleavage of Disulfide Bonds in Intermolecularly-Linked Polypeptide Ions. Analytical chemistry, 88(18), 8972-8979.

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Immunofluorescence Staining of Cell Organelles

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Bafilomycin A1, epinephrine, nifedipine and somatostatin were purchased from Sigma (Buchs, Switzerland). Calibration beads were from Molecular Probes (Zug, Switzerland). The antibodies against GFP, Insulin and Synaptophysin were from Chemicon (Zug, Switzerland), Abcam (Cambridge, UK) and Sigma (Buchs, Switzerland), respectively. All other chemicals (unless indicated) were obtained from Invitrogen (Basel, Switzerland).

Bergeron A., Pucci L., Bezzi P, & Regazzi R. (2014). Analysis of Synaptic-Like Microvesicle Exocytosis of B-Cells Using a Live Imaging Technique. PLoS ONE, 9(2), e87758.

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Isolation and Culture of Thyroid Cancer Cells

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Cancer tissues obtained from mPTC^GFP were minced in lysis buffer (Collagenase I (Sigma) 1 mg/ml; Dispase II (Invitrogen) 0.5 mg/ml in PBS) and shaked on an orbital shaker at 37 °C for 60 min. Dissociated tumor cells were filtered with 40 μm cell strainer (#352340, BD Falcon) and centrifugated at 500 g for 5 min, the pellet was resuspended in cold Ham’s F-12 Nutrient Mix (#11765062, Gibco). The isolation of tumor cells was next performed on FACSAria Cell Sorter (Becton Dickinson) according to GFP signal, and positive cells were washed with PBS and then cultured in F-12 medium supplemented with 10% FBS, 1% penicillin/streptomycin, 5 mg/L transferrin (Sigma-Aldrich), 10 mg/L bovine insulin (Solarbio), 3.5 mg/L hydrocortisone (Sigma-Aldrich), 10 mg/L somatostatin (Sigma-Aldrich), 0.02 mg/L Gly-His-Lysacetate (Sigma-Aldrich) and 1 IU/L bovine thyroid stimulating hormone (TSH) (Sigma-Aldrich) related to the 6H medium at 37 °C⁵⁹. Next, Primary cells treated with 10 μM PLX4032 or 2 μM SCH772984 were subjected to western blot and RT-qPCR analysis. Primary cells from mPTC mice were directly cultured in complete F-12 medium. Thyrocytes were confirmed by immunofluorescence staining of thyroglobulin and representative photos were taken by a fluorescence microscopy.

Zhang P., Guan H., Yuan S., Cheng H., Zheng J., Zhang Z., Liu Y., Yu Y., Meng Z., Zheng X, & Zhao L. (2022). Targeting myeloid derived suppressor cells reverts immune suppression and sensitizes BRAF-mutant papillary thyroid cancer to MAPK inhibitors. Nature Communications, 13, 1588.

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FRTL-5 Cell Culture Protocol

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FRTL-5 cells were kindly provided by Dr. Zheng Xuqin (Department of Endocrinology, First Affiliated Hospital of Nanjing Medical University) [28 (link)], who was kindly gifted these cells by Professor Michael Derwahl (Department of Medicine, St. Hedwig Hospital) [29 (link)]. Coon’s modified Ham’s F-12 medium, transferrin, bovine insulin, hydrocortisone, somatostatin, glycyl-L-histidyl-L-lysine acetate, and bovine TSH were purchased from Sigma-Aldrich (USA). Newborn calf serum was purchased from Gibco (USA). Penicillin and streptomycin were purchased from HyClone (USA). PCB118 (purity: 100%, analyzed using GC/MS) was purchased from AccuStandard (USA) and was dissolved initially in 612 μL dimethyl sulfoxide (DMSO) and maintained as a 25 mM stock solution in the dark. When needed, this solution was diluted to the required concentration with culture media. The final concentration of DMSO in culture medium was maintained below 0.1%.

Yang H., Chen H., Guo H., Li W., Tang J., Xu B., Sun M., Ding G., Jiang L., Cui D., Zheng X, & Duan Y. (2015). Molecular Mechanisms of 2, 3′, 4, 4′, 5-Pentachlorobiphenyl-Induced Thyroid Dysfunction in FRTL-5 Cells. PLoS ONE, 10(3), e0120133.

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Cultivation of Rat Thyroid Cells

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The differentiated strain of rat-thyroid cells (Fisher-rat-thyroid-line-5; FRTL5; American Type Culture Collection, Manassas, VA; ATCC CRL 8305, F1 subclone) served as an in vitro experimental substrate. Cells were grown for 1 week in 6H medium (Coon’s Modified Ham’s F-12 Medium (Sigma Chemical Co.) supplemented with 5% adult calf serum (BioWhittaker, Inc., Walkersville, MD) and a six-hormone mixture containing insulin, somatostatin, hydrocortisone, transferrin, glycyl-histidyl-lysine, and TSH (Sigma Chemical Co.).

Coperchini F., Croce L., Pignatti P., Ricci G., Gangemi D., Magri F., Imbriani M., Rotondi M, & Chiovato L. (2020). The new generation PFAS C6O4 does not produce adverse effects on thyroid cells in vitro. Journal of Endocrinological Investigation, 44(8), 1625-1635.

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FRTL-5 Thyroid Cell Culture Protocol

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Fischer rat thyroid line cells (FRTL-5, American Type Culture Collection, Rockville, MD, CRL 8305) derived from the normal thyroid gland of Fischer rats were maintained in a 37 °C humidified incubator supplied with 95%/5% air/CO₂. FRTL-5 cells were cultured according to the supplier's recommendations in Ham’s F12K medium with 2 mM l-glutamine and adjusted to contain 1.5 g/L sodium bicarbonate. 0.5% bovine calf serum and six additional hormones required for FRTL-5 cells to proliferate and maintain thyroid function were added to the medium, as specified by the originators of this cell line [22] , [23] . These six hormones included 1 mU/mL thyroid stimulating hormone (TSH), 0.01 mg/mL insulin, 10 nM hydrocortisone, 0.005 mg/mL transferrin, 10 ng/mL somatostatin, and 10 ng/mL glycyl-l-histidyl-l-lysine acetate (All reagents from Sigma, St Louis, MO) [24] (link). Growth medium was used for cell expansion and to acclimate the cells prior to initiating the experiments. For the experiments, growth medium was modified so that it either contained 10 mU/mL TSH or no TSH as indicated below. Cells were passaged at 70% confluency and harvested at passages four through eight for the experiments.

Wagner A.P., Chinnathambi S., Titze I.R, & Sander E.A. (2016). Vibratory stimulation enhances thyroid epithelial cell function. Biochemistry and Biophysics Reports, 8, 376-381.

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Intraperitoneal Glucose Tolerance Test in Mice

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For intraperitoneal glucose tolerance tests, mice were intraperitoneally administered 1 g/kg body weight glucose for HFD-fed mice and 2 g/kg body weight glucose for NC-fed mice, with or without continuous intravenous administration of somatostatin (3 μg/kg/min) (Sigma) through a jugular catheter from 1 h before glucose administration. Blood glucose levels were measured using a GLUCOCARD G+ Meter (Arkray, Kyoto, Japan) and a commercial colorimetric kit (Wako). Plasma insulin levels were measured using a Mouse Insulin ELISA Kit (Shibayagi, Gunma, Japan).

Watanabe H., Inaba Y., Kimura K., Matsumoto M., Kaneko S., Kasuga M, & Inoue H. (2018). Sirt2 facilitates hepatic glucose uptake by deacetylating glucokinase regulatory protein. Nature Communications, 9, 30.

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Pancreatic Substrate Clamp in Pigs

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On d 5, after overnight fasting, animals were placed in a sling restraint system to perform pancreatic-substrate clamps for 2 h, as previously described (5 (link),11 (link)). The clamp was initiated by infusing 100 μg/kg/h of somatostatin (Bachem, Torrance, CA). Glucose and AAs were also clamped ± 10th percentile of the individual baseline fasting levels by monitoring their concentration every 5 min and adjusting the infusion rate of 10% dextrose (~60–80 mg/dl) and a balanced mixture of essential and nonessential AAs (~500 nmol BCAA/ml) (5 (link),11 (link)).
Fifteen min after starting the somatostatin infusion, replacement glucagon (150 ng/kg/h; Sigma-Aldrich, Milwaukee, WI) and insulin at either 7 ng/kg^0.66/min to achieve plasma concentrations of 2–5 μU/ml to simulate a fasting insulinemic state (CON, n = 4; CLP, n = 4), or at 40 ng/kg^0.66/min (CON + I, n = 5; CLP + I, n = 5) to replicate fed levels of insulin (~30 μU/ml) observed in healthy neonatal pigs, were also infused.

Manjarín R., Suryawan A., Koo S.J., Wilson F.A., Nguyen H.V., Davis T.A, & Orellana R.A. (2016). Insulin modulates energy and substrate sensing and protein catabolism induced by chronic peritonitis in skeletal muscle of neonatal pigs. Pediatric research, 80(5), 744-752.

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