S5261

Manufactured by Merck Group

Sourced in Japan, United States

The S5261 is a laboratory equipment product. It is designed for use in scientific research and analysis. The core function of the S5261 is to provide a controlled and consistent environment for various experimental procedures.

Automatically generated - may contain errors

Lab products found in correlation

Transferrin, by Merck Group (5 mentions) Ascorbic acid, by Fujifilm (3 mentions) H0888, by Merck Group (3 mentions) E9644, by Merck Group (2 mentions) P3932, by Merck Group (2 mentions) I6634, by Merck Group (2 mentions) E2758, by Merck Group (2 mentions) L6520, by Merck Group (2 mentions) I4011, by Merck Group (1 mentions) Epidermal growth factor (egf), by Merck Group (1 mentions) F1141, by Merck Group (1 mentions) I0516, by Merck Group (1 mentions) Hydrocortisone, by Merck Group (1 mentions) Insulin, by Thermo Fisher Scientific (1 mentions) Oxldl, by Thermo Fisher Scientific (1 mentions) J65591, by Thermo Fisher Scientific (1 mentions) F 12k medium, by American Type Culture Collection (1 mentions) T3309, by Merck Group (1 mentions) At140, by HiMedia (1 mentions) C1386, by Merck Group (1 mentions)

15 protocols using s5261

Luteinizing and Non-Luteinizing Granulosa Cell Culture

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To prepare luteinizing and non-luteinizing GCs, the culture medium was replaced with a fresh medium containing 0.1% of BSA, 5 ng/ml sodium selenite
(Sigma-Aldrich; S5261), 5 µg/ml transferrin (Sigma-Aldrich; T4132), and 0.5 mM ascorbic acid (Wako-Pure Chemical Industries Osaka, Japan; 031-12061), and the
cells were then incubated in a normal culture atmosphere (20% O₂, 5% CO₂, and 75% N₂) with or without insulin (2 µg/ml;
Sigma-Aldrich; I4011) in the medium in combination with forskolin (10 µM; Research Biochemicals International, Natick, MA, USA; 70-0501-05) for 24 h. Insulin
and insulin-like growth factor I (IGF-I) are known to stimulate proliferation of (and P4 production in) GCs [27 (link),28 (link),29 (link),30 (link),31 (link)]. In addition, forskolin increases intracellular cyclic AMP concentration via activation of adenylate cyclase [32 (link)]. Insulin in combination with forskolin mimics the effects of luteinizing hormone (LH) and activates adenylate cyclase via upregulation of P4
[33 (link)]. The concentration of insulin and forskolin was selected according to other reports [26 (link), 34 ].

F.A.D.H.I.L.L.A.H., YOSHIOKA S., NISHIMURA R., YAMAMOTO Y., KIMURA K, & OKUDA K. (2016). Hypoxia-inducible factor 1 mediates hypoxia-enhanced synthesis of progesterone during luteinization of granulosa cells. The Journal of Reproduction and Development, 63(1), 75-85.

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Evaluating Biofilm Eradication Concentrations on Voice Prostheses

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We determined the minimum mature BEC on voice prostheses of sodium selenite (Sigma‒Aldrich, S5261) as described previously.22 (link) Briefly, the medical silicone membrane was placed in a 24-well plate, and 1 mL of mixed-bacteria solution was added to each well and cultured at 37°C for 24 hours. Then, different concentrations of sodium selenite (0, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, 1.00 mg/mL) were added to the various wells, and the plate was cultured at 37°C for 24 hours. The number of colonies in each well was evaluated by the plate counting method. By comparison with the negative control (sodium selenite concentration: 0 mg/mL), the lowest sodium selenite concentration resulting in a 50% reduction in the number of colonies on the voice prosthesis biofilms was taken as the BEC₅₀, the lowest sodium selenite concentration resulting in a 70% reduction in the number of colonies was the BEC₇₀, and the lowest sodium selenite concentration resulting in a 90% reduction in the number of colonies was the BEC₉₀.

Zhang Y., Niu Y., Huo H., Wang J., Jin X, & Yang H. (2022). Inhibition and Removal of Mature Mixed-Bacteria Biofilms on Voice Prostheses by Sodium Selenite. Infection and Drug Resistance, 15, 7799-7810.

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Genetic Screening of E3 Ligase Knockdowns

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HeLa cells were transduced with three lentiviral shRNA pools containing 2,833 shRNAs against 616 E3 ligase genes or NS shRNA in triplicate at 0.2 multiplicity of infection (MOI) to prevent superinfection and to ensure that each cell received no more than one shRNA. After infection, HeLa cells were selected with puromycin (0.2 μg ml⁻¹) for 7 days to enrich for HeLa cells expressing shRNA. After puromycin selection, HeLa cells were grown in serum-free DMEM containing trace elements (D0547, Sigma) supplemented with 50 ng ml⁻¹ EGF (E9644, Sigma), 20 ng ml⁻¹ FGF (SRP3043, Sigma), 100 nM hydrocortisone (H0888, Sigma), 0.5 μg ml⁻¹ transferrin (T1147, Sigma), 5 ng ml⁻¹ selenium (S5261, Sigma), 0.5 μg ml⁻¹ fibronectin (F1141, Sigma) and 0.05 μg ml⁻¹ insulin (I0516, Sigma). Media was changed every 3 days and cells were split at a 1:4 ratio every 7 days. After four passages all cells carrying NS and shRNAs from pool 2 were dead. Surviving cells from pools 1 and 3 were collected and genomic DNA was isolated. The integrated shRNAs were PCR-amplified using primers specific to the shRNA vector (pLKO.1) and listed in Supplementary Table 3. Samples were sequenced using primer SP6 (Supplementary Table 3) to identify candidate shRNAs.

Nagarajan A., Petersen M.C., Nasiri A.R., Butrico G., Fung A., Ruan H.B., Kursawe R., Caprio S., Thibodeau J., Bourgeois-Daigneault M.C., Sun L., Gao G., Bhanot S., Jurczak M.J., Green M.R., Shulman G.I, & Wajapeyee N. (2016). MARCH1 regulates insulin sensitivity by controlling cell surface insulin receptor levels. Nature Communications, 7, 12639.

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Culturing Human Aortic Smooth Muscle Cells

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Human aortic vascular smooth muscle cells (VSMCs) were purchased from American Type Culture Collection (T/G HA-VSMC, CRL-1999™, Manassas, VA), and were cultured according to the supplier's instructions at 37°C, 5% CO₂. To make the complete growth medium, the following components (final concentrations) were added to the F-12K base medium (30-2004, ATCC): 0.05 mg/ml ascorbic acid; 0.01 mg/ml insulin (12585014, Gibco); 0.01 mg/ml transferrin (T8158, Sigma, St. Louis, MO); 10 ng/ml sodium selenite (S5261, Sigma) 20% fetal bovine serum; 10 mM HEPES (7365-45-9, American Bioanalytical, Natick, Massachusetts); 10mM TES (T5691, Sigma).
VSMC in passages 2–6 were used for all experiments. Controls were treated with medium alone plus vehicle where applicable and maintained for the duration of all experiments. Treatment medium consisted of basal medium (F-12K Medium, 30-2004, ATCC) supplemented with 100 ng/mL rWNT5A or rWNT3A (human recombinant WNT5A & WNT3A, 645-WN & 5036-WN respectively, R&D Systems, Minneapolis, MN), 10 ng/mL platelet-derived growth factor-BB (PDGF-BB, GF149, Millipore Sigma, Burlington, MA), or 100 μg/mL low-density lipoproteins (n-LDL and ox-LDL, J65039 & J65591, Alfa Aesar) for the indicated duration. Blocking experiments were performed according to the methods described above.

Ackers I., Szymanski C., Silver M.J, & Malgor R. (2020). Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line. Frontiers in Cardiovascular Medicine, 7, 567837.

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3D Spheroid Culture of Mammary Cells

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Spheroids were cultured in 3% polyHEMA-coated (P3932, Sigma-Aldrich) dishes in basal medium supplemented with 250 ng/mL insulin (I6634, Sigma-Aldrich), 0.5 μg/mL hydrocortisone (H0888, Sigma-Aldrich), 2.6 ng/mL sodium selenite (S5261, Sigma-Aldrich), 10 μg/mL transferrin (T3309, Sigma-Aldrich), 27.3 pg/mL estradiol (E2758, Sigma-Aldrich) and 5 μg/mL prolactin (L6520, Sigma-Aldrich). These were maintained in culture for 1 to 7 days. Spheroids were visualized using the Olympus IX73 fluorescence microscope.

Arora G., Banerjee M., Langthasa J., Bhat R, & Chatterjee S. (2023). Targeting metabolic fluxes reverts metastatic transitions in ovarian cancer. iScience, 26(11), 108081.

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Spheroid Culture in Poly-HEMA Coated Dishes

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Spheroids were cultured in tissue culture dishes coated with 3% poly-2-hydroxyethyl methacrylate (polyHEMA) (P3932; Sigma-Aldrich) solution. Culture dishes were coated overnight under sterile conditions. PolyHEMA solution was prepared in 95% absolute ethanol. PolyHEMA coating prevented cell attachment and allowed spheroid formation in suspension. Cells were seeded according to the requirement for experiment in defined medium: DMEM: F12 (1:1) (HiMedia AT140) supplemented with 0.5 µg/ml hydrocortisone (Sigma-Aldrich, H0888), 250 ng/ml insulin (Sigma-Aldrich, I6634), 2.6 ng/ml sodium selenite (Sigma-Aldrich, S5261)), 27.3 pg/ml estradiol (Sigma-Aldrich, E2758), 5 µg/ml prolactin (Sigma-Aldrich L6520, 10 µg/ml transferrin (Sigma-Aldrich, T3309). Spheroids were visualized using phase contrast or bright-field microscopy and collected from the cultures by centrifugation at 0.1–0.2g for 5 min.

Langthasa J., Sarkar P., Narayanan S., Bhagat R., Vadaparty A, & Bhat R. (2021). Extracellular matrix mediates moruloid-blastuloid morphodynamics in malignant ovarian spheroids. Life Science Alliance, 4(10), e202000942.

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Granulosa Cell Progesterone Production

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To prepare luteinizing and non-luteinizing granulosa cells, the culture medium was replaced with fresh medium containing 0.1%
BSA, 5 ng/ml sodium selenite (Sigma-Aldrich; S5261), 5 µg/ml transferrin (Sigma-Aldrich; T4132) and 0.5 mM ascorbic acid
(Wako-Pure Chemical Industries, Osaka, Japan; 031–12061), and the cells were then incubated under a normal culture atmosphere
(20% O₂, 5% CO₂, 75% N₂) without or with insulin (2 µg/ml; Sigma-Aldrich; I4011), forskolin
(10 µM; Research Biochemicals International, Natick, MA, USA; 70–0501-05) or insulin (2 µg/ml) in combination with forskolin
(10 µM) for 24 h. The concentration of insulin and forskolin was selected based on a previous report [25 ]. The culture media from these cultured cells were collected to determine the effect of insulin and
forskolin treatment on P4 production for 24 h.
To determine the effect of hypoxia on P4 production, mRNA and protein expressions of STAR, CYP11A1 and HSD3B, the luteinizing
and non-luteinizing granulosa cells were incubated under various O₂ concentrations, 20% O₂ (normoxia),
10% O₂ (hypoxia) or 5% O₂ (hypoxia), for 24 h in small individual culture chambers. The chambers were
refilled with a nonstandard gas mixture, as described previously [10 (link)], containing the
indicated O₂ level (20%, 10% or 5% O₂) and 5% CO₂ in an N₂ base.

F.A.D.H.I.L.L.A.H., YOSHIOKA S., NISHIMURA R, & OKUDA K. (2014). Hypoxia Promotes Progesterone Synthesis During Luteinization in Bovine Granulosa Cells. The Journal of Reproduction and Development, 60(3), 194-201.

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Cytotoxicity Assay of iAs, Selenite, and Curcumin

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EpASCs from neonates born to mothers in control group were employed for the assay. Cells were seeded (2*10⁴ cells/well) in 96-well plates and allowed to adhere for 24 hours in a CO₂ incubator. iAs and selenite were dissolved in MilliQ water; curcumin was dissolved in DMSO. Final concentration of DMSO in the assay was ≤0.05%; flasks from the control group received only the vehicle. Cells were exposed (in triplicate, for 24 hours) to serial dilutions (40 to 1.25 μM) of arsenite (NaAsO₂, Sigma catalogue #S7400) or selenite (Na₂SeO₃, Sigma catalogue #S5261) or curcumin (Sigma catalogue #C1386). After 24h exposure, MTT (10μl of 5mg/ml) was added and cells were further incubated for 3-4hrs. The formazan crystals formed in viable cells were dissolved in DMSO. A₅₄₀ was determined using a microplate reader (Microquant, Bio-Tek, USA) to establish EC₅₀ value of arsenite, selenite and curcumin (S1a Fig).

Poojan S., Kumar S., Verma V., Dhasmana A., Lohani M, & Verma M.K. (2015). Disruption of Skin Stem Cell Homeostasis following Transplacental Arsenicosis; Alleviation by Combined Intake of Selenium and Curcumin. PLoS ONE, 10(12), e0142818.

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Culturing human colonic epithelial and fibroblast cells

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The human colonic epithelial cell lines 1CT were previously described (25 (link)). 1CT cells were cultured in complete medium, composed as follows: high‐glucose Dulbecco’s Modified Eagle (DMEM) medium/medium 199 (4530, Sigma‐Aldrich Merck, Darmstadt, Germany, at ratio 4:1), supplemented with 2% fetal bovine serum (FBS; 10270-106, Gibco ThermoFisher Scientific, Waltham, MA, USA), epidermal growth factor (EGF; 20 ng/ml, E9644, Sigma‐Aldrich Merck), hydrocortisone (1 mg/ml, H0396, Sigma‐Aldrich Merck), insulin (10 mg/ml, I9278, Sigma‐Aldrich), transferrin (2 mg/ml, T0665, Sigma‐Aldrich Merck), sodium selenite (5 nM, S5261, Sigma‐Aldrich Merck), and geneticin (G418 sulfate) sulfate (50 μg/ml, G1397, Sigma‐Aldrich Merck).
Human colonic fibroblasts (CRL-1459, ATCC, Manassas, VA, USA) were maintained in Eagle’s Minimum Essential Medium (EMEM; 30-2003, ATCC, Manassas, VA, USA) supplemented with 10% of FBS (10270-106, Gibco, ThermoFisher Scientific) and 10 μg/ml of ciprofloxacin (17850, Sigma‐Aldrich Merck).

Lopez Chiloeches M., Bergonzini A., Martin O.C., Bergstein N., Erttmann S.F., Aung K.M., Gekara N.O., Avila Cariño J.F., Pateras I.S, & Frisan T. (2024). Genotoxin-producing Salmonella enterica induces tissue-specific types of DNA damage and DNA damage response outcomes. Frontiers in Immunology, 14, 1270449.

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Isolation and Culture of Mouse Spermatogonial Stem Cells

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Mouse SSCs were isolated from the testis of six to seven days postpartum (dpp) C57BL/6JNarl or SMA-like mice [C57BL/6/Tg(SMN2) Hung Smn1^{tm1 Hung}] (National Laboratory Animal Center, NLAC, Taipei, Taiwan) [33 (link)]. After Collagenase IV and Trypsin digestion, testicle cells were incubated with anti-CD90.2 (THY1) antibody and then labeled with anti-biotin microbeads (130-090-485, Miltenyi Biotec, Bergisch Gladbach, Germany). 2 × 10⁵ SSCs were cultured in gelatin-coated 24-well plate seed with mitomycin C treated mouse embryonic fibroblast (MEF) feeders and maintained in SSC medium composed of complete StemPro-34 medium (10639011) supplemented with 5 mg/mL of BSA (A4378, Sigma), 1% of ES cell-qualified heat-inactivated FBS (16000-044), D-(+)-glucose (6 mg/mL, G6152, Sigma), 2-mercaptoethanol, ITS, P/S, GlutaMax, NEAA, MEM vitamin solution (11120052), 0.14% (wt/vol) of sodium bicarbonate (S5761, Sigma), sodium selenite (30 nM, S5261, Sigma), sodium pyruvate (30 μg/mL, P3662, Sigma), Putrescine (60 μM, P7630, Sigma), ascorbic acid (100 nM, A4403, Sigma), D-biotin (10 μg/mL, B4639, Sigma), progesterone (60 ng/mL, P0130, Sigma), β-estradiol (30 ng/mL, E88775, Sigma), EGF (20 ng/mL), GDNF (10 ng/mL), and bFGF (10 ng/mL). Half of the medium were changed daily till confluence and passed by TrypLE to new feeder for further culture.

Chang W.F., Peng M., Hsu J., Xu J., Cho H.C., Hsieh-Li H.M., Liu J.L., Lu C.H, & Sung L.Y. (2021). Effects of Survival Motor Neuron Protein on Germ Cell Development in Mouse and Human. International Journal of Molecular Sciences, 22(2), 661.

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