Sd 208

Manufactured by Merck Group

Sourced in United States

The SD-208 is a laboratory equipment product manufactured by Merck Group. It serves as a tool for performing various scientific experiments and analyses. The core function of the SD-208 is to provide a controlled environment for conducting research and testing.

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Lab products found in correlation

Sb431542, by Merck Group (2 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Stem cell factor (scf), by Thermo Fisher Scientific (1 mentions) Acetaldehyde, by Merck Group (1 mentions) Stemspan sfem, by STEMCELL (1 mentions) Methocult gf m3434 methylcellulose, by STEMCELL (1 mentions) Cilengitide, by Selleck Chemicals (1 mentions) Gant58, by Santa Cruz Biotechnology (1 mentions) High glucose dmem, by Corning (1 mentions) Ly364947, by Merck Group (1 mentions) Thiazovivin, by Merck Group (1 mentions) A83 01, by Bio-Techne (1 mentions) Gw788388, by Merck Group (1 mentions) Recombinant mouse tgf β1, by R&D Systems (1 mentions) Sr 3677, by Merck Group (1 mentions) Y 27632, by Enzo Life Sciences (1 mentions) Resomer rg 503h, by Merck Group (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti mmp2, by Cell Signaling Technology (1 mentions) Anti phosphorylated smad2, by Cell Signaling Technology (1 mentions)

17 protocols using sd 208

Hematopoietic Stem Cell Assays

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For clonogenic assays,20,000 Lin⁻ cells per condition (in triplicate) and 2000 sorted LSK cells per condition (in triplicate) were plated in Methocult GF M3434 methylcellulose (03444,Stem Cell Technologies) and cultured for 7 days. The TGFβ inhibitors such as AVID200 (Forbius), SD208 (S7071, Sigma-Aldrich) and 1D11 (Genzyme) were added in graded concentrations in the cultures. For acetaldehyde treatments, bone marrow cells were exposed to increasing doses of acetaldehyde (00070–100ML, Sigma-Aldrich) for 4 h, washed and then cultured in Methocult GF M3434 methylcellulose. The hematopoietic colonies were scored after 7 days of culture at 37 °C and 5% CO₂.
For proliferation assays, sorted LSK cells were cultured in serum-free medium StemSpan SFEM (09600, StemCell Technologies) containing 1% penicillin/streptomycin (15140–122, GIBCO), 2% L-glutamine (25030–081GIBCO), 100 ng/ml TPO (315–14-10UG, Peprotech) and 100 ng/ml SCF (250–03-10UG, Peprotech). After 48 h in culture, DNA damage was assessed by immunofluorescence staining the cells to detect γH2AX foci formation (2577s, Cell Signaling) as described [13 (link)].

Rodríguez A., Yang C., Furutani E., de Teresa B.G., Velazquez M., Filiatrault J., Sambel L.A., Phan T., Flores-Guzmán P., Sánchez S., Orozco A.M., Mayani H., Bolukbasi O.V., Färkkilä A., Epperly M., Greenberger J., Shimamura A., Frías S., Grompe M., Parmar K, & D’Andrea A.D. (2020). Inhibition of TGFβ1 and TGFβ3 promotes hematopoiesis in Fanconi anemia. Experimental hematology, 93, 70-84.e4.

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Cytotoxicity of Targeted Inhibitors

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2D films or 3D scaffolds were seeded with tumor cells, cultured for 24 h, and then treated with 10 μM Cilengitide (Selleck Chemicals, Huston, TX, USA), 10 μM GANT 58 (Santa Cruz Biotechnology, Dallas, TX, USA), 100 nM SD208 (Sigma-Aldrich, St. Louis, MO, USA), or DMSO as a control in serum-free media. After 48 h of drug treatment, RNA was harvested for qRT-PCR analysis.

Dadwal U.C., Merkel A.R., Page J.M., Kwakwa K.A., Kessler M, & Rhoades J.A. (2020). 3D Bone Morphology Alters Gene Expression, Motility, and Drug Responses in Bone Metastatic Tumor Cells. International Journal of Molecular Sciences, 21(18), 6913.

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Culturing Human Cardiac Fibroblasts

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Human cardiac fibroblasts (Passage number 4–6) (Cell Applications, San Diego, CA, USA) were kept in culture using DMEM High Glucose (Corning, Somerville, NY, USA) supplemented with fetal bovine serum (FBS, 10%) (Hyclone, South Logan, UT, USA), penicillin-streptomycin (P/S, 1%) (Gibco, Waltham, MA, USA), and SD208 (3 µM, TGF-β pathway inhibitor) (Sigma-Aldrich, St. Louis, MO, USA) at 37 °C with 5% CO₂. The culture was maintained with daily half-media changes until they reached approximately 80% confluency.

Basara G., Ozcebe S.G., Ellis B.W, & Zorlutuna P. (2021). Tunable Human Myocardium Derived Decellularized Extracellular Matrix for 3D Bioprinting and Cardiac Tissue Engineering. Gels, 7(2), 70.

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Stem Cell Differentiation Regulator Compounds

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Recombinant peptide and small molecules are as follows: A83-01 (Tocris 2939), Y-27632 (Enzo Life Science ALX-270-333-M005), Recombinant Mouse TGF-β 1 (Novoprotein CA59, R&D Systems 7666-MB-005), GW788388 (Sigma-SML0116-5 mg), LY-364947 (Sigma-L6293-5MG), SD-208 (Sigma-S7071-5MG), Thiazovivin (Sigma-SML1045-5 mg) and SR-3677 (Sigma-SML0774-5 mg).

Zhao Y., Londono P., Cao Y., Sharpe E.J., Proenza C., O'Rourke R., Jones K.L., Jeong M.Y., Walker L.A., Buttrick P.M., McKinsey T.A, & Song K. (2015). High-efficiency reprogramming of fibroblasts into cardiomyocytes requires suppression of pro-fibrotic signalling. Nature Communications, 6, 8243.

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Preparation and Characterization of Membrane-Coated PLGA Nanoparticles

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PLGA nanoparticles were prepared with o/w single emulsion. 1mL of PLGA (resomer® RG 503 H, Sigma Aldrich, USA) dissolved in dichloromethane (6 mg/ml) was added dropwise into 6ml of 0.5% PVA solution. SD-208 (Sigma Aldrich, USA) was mixed with PLGA solution with a polymer-to-drug ratio of 10:1. For the fluorescence imaging experiment, Cy5.5 NHS or DiD (Sigma Aldrich, USA) was loaded into the PLGA solution at 5% (w/w). The mixture was then agitated with 200RPM until the dichloromethane was evaporated entirely. After the dichloromethane was evaporated, the solution was centrifuged and washed twice with distilled water at 20,000 g for 20 min to remove the unloaded-free drugs or dyes Membrane-coated nanoparticles were prepared with the sonication method. In short, the cell membrane was mixed with PLGA NP with a 1:1 weight ratio (w/w) and sonicated in a water bath sonicator for 3 min. In order to remove the uncoated nanoparticles, the mixture was centrifuged at 20,000 g for 20 min.

Kim J., Kim M., Yong S.B., Han H., Kang S., Lahiji S.F., Kim S., Hong J., Seo Y, & Kim Y.H. (2023). Engineering TGF-β inhibitor-encapsulated macrophage-inspired multi-functional nanoparticles for combination cancer immunotherapy. Biomaterials Research, 27, 136.

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Recombinant TGF Treatment Protocol

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Recombinant TGF (R&D) was resuspended in phosphate-buffered saline (PBS) and added to cells at the desired concentrations. LDN 193189 (LDN; Sigma), SB431542 (Sigma), and SD208 (Sigma) were reconstituted using the company directions and at the desired concentrations. These recommended concentrations of the drugs were found not to be toxic to the cells as determined by trypan blue exclusion.

Poole E., Carlan da Silva M.C., Huang C., Perera M., Jackson S., Groves I.J., Wills M., Rana A, & Sinclair J. (2021). A BMPR2/YY1 Signaling Axis Is Required for Human Cytomegalovirus Latency in Undifferentiated Myeloid Cells. mBio, 12(3), e00227-21.

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Inhibition of TGF-β Signaling Pathway

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Human recombinant TGF-β1 was obtained from R&D Systems (Minneapolis, MN). The TGF-β RI (TβRI) kinase inhibitor SB431542 was obtained from Millipore (Billerica, MA), and SD-208 was purchased from Sigma (St Louis, MO). The following antibodies were used in this study: anti-PPM1A, anti-MMP2, anti-MMP9, anti-Smad2, anti-Smad3, anti-phosphorylated Smad2 and anti-phosphorylated Smad3 (Cell Signaling Technology, Beverly, MA) as well as anti-E-cadherin, anti-GAPDH, anti-CK19, anti-phosphorylated Smad2/3 and anti-Vim (Santa Cruz, CA, USA). SB431542 was applied during the TGF-β1 perfusion.

Geng J., Fan J., Ouyang Q., Zhang X., Zhang X., Yu J., Xu Z., Li Q., Yao X., Liu X, & Zheng J. (2014). Loss of PPM1A expression enhances invasion and the epithelial-to-mesenchymal transition in bladder cancer by activating the TGF-β/Smad signaling pathway. Oncotarget, 5(14), 5700-5711.

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Neonatal Rat Ventricular Myocyte Isolation and Treatment

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Neonatal rat ventricular myocytes (NRVMs) were isolated from Sprague Dawley rats (50–60 rats pooled, male and female, 1 day old), kept on ice before isolation, and isolated using previously identified method.³⁰ NRVMs were cultured for 24 hours in growth media, rinsed, and cultured 24 hours in serum‐free media (minimum essential media (MEM), HEPES, B12, Bromodeoxyuridine (BrdU), BSA, insulin, transferrin) to prevent growth of fibroblasts. CM from fibroblasts was used to treat NRVMs for 48 hours before measuring cell size. SD208 (Sigma Aldrich S7071) was used in combination with CF CM treatment of NRVMs at 3μmol/L. The same protocol was used for insulin growth factor 1 (IGF‐1) and osteopontin treatments. IGF‐1 (R&D Systems 4326‐RG‐050) and osteopontin (R&D Systems 6359‐OP‐050) were added NRVMs in serum‐free media and cultured for 48 hours before NRVMs were trypsinized and cell size was measured by Coulter Counter.

Ceccato T.L., Starbuck R.B., Hall J.K., Walker C.J., Brown T.E., Killgore J.P., Anseth K.S, & Leinwand L.A. (2020). Defining the Cardiac Fibroblast Secretome in a Fibrotic Microenvironment. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 9(19), e017025.

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Cardiac Fibroblast Isolation and Phenotype Modulation

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Fbs from the LV were isolated by a chunk enzymatic digestion protocol as described before^{20 (link)}. After first passage, Fbs were seeded at a density of 3000 cells/cm² and cultured in DMEM (ThermoFisher) medium with 10% foetal bovine serum (FBS) and 1% of penicillin/streptomycin (Invitrogen). To induce specific Fb phenotypes, cells were either treated for 6 days with recombinant human TGF-β1 (400 pmol/L; PeproTech) or with SD-208 (3 μmol/L; Sigma-Aldrich), a specific TGF-β-receptor-I (TGF-β-RI) kinase inhibitor to respectively promote and suppress differentiation. After 6 days, the phenotype was assessed by protein expression and morphology, and functionally as the capacity for proliferation, migration, and contraction of a floating gel. These pre-treated Fbs and MyoFbs were then processed for co-culture with freshly isolated CMs. The supplemental material and Figures S1 detail the preparation and properties of the fibroblastic cells and the subsequent setup of the co-cultures.

Nagaraju C.K., Dries E., Gilbert G., Abdesselem M., Wang N., Amoni M., Driesen R.B, & Sipido K.R. (2019). Myofibroblast modulation of cardiac myocyte structure and function. Scientific Reports, 9, 8879.

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Prostate Cancer Cell Line Assays

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PC-3, DU-145 and RM-1 prostate cancer cells were obtained from the American Type Culture Collection. The cancer cells were cultured in RPMI 1640 supplemented with 10% FBS. DTX (MedChemExpress, Monmouth Junction, New Jersey, USA), which promotes microtubule stabilization and inhibits the mitosis of cancer cells; cisplatin (MedChemExpress), a replicative polymerase inhibitor, were used at concentrations of 0, 25, 50, 100 nM and 0, 10, 20, 30 µM, respectively. Paclitaxel (PTX, MedChemExpress), was used at concentration of 200 nM. Additionally, indicated concentrations of the ATM inhibitor KU55933 (Sigma-Aldrich, St. Louis, Missouri, USA) or the NEMO binding peptide (NBD) (Enzo Biochem, New York, USA) were added at 60 min before the induction of PD-L1. For inhibitor screening, S3I-201(100 µM), Fludarabine (10 µM), UO126-EtOH (10 µM), Bay11-7082 (10 µM), JSH-23 (5 µM), SD-208 (2 µM) which obtained from Sigma-Aldrich were added and the cells were collected after 6 hours.

Wang Z., Zhang X., Li W., Su Q., Huang Z., Zhang X., Chen H., Mo C., Huang B., Ou W., Chen J., Zhao G., Chen L, & Shao L. (2021). ATM/NEMO signaling modulates the expression of PD-L1 following docetaxel chemotherapy in prostate cancer. Journal for Immunotherapy of Cancer, 9(7), e001758.

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