Reagent-grade chemicals were mostly obtained from Sigma and Himedia. Heat-inactivated fetal bovine serum (FBS, Himedia), Dulbecco’s Modified Eagle’s Medium F12/ (DMEM/F12, Himedia). 1% Penicillin-Streptomycin (Sigma), MTT [3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyl tetrazolium bromide] were procured from Sigma-Aldrich Chemicals (St. Louis, MO). 0.25% Trypsin-EDTA 1X Solution (Himedia), DCFH-DA (Sigma) and other routine chemicals were purchased from Sigma/Merck.
Trypsin edta solution 1x
Manufactured by HiMedia
Trypsin-EDTA Solution 1x is a cell dissociation reagent used in cell culture applications. It contains the enzyme trypsin and the chelating agent EDTA, which work together to break down the proteins that hold cells together, allowing them to be easily dispersed and passaged. The solution is provided at a 1x concentration, ready-to-use.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by HiMedia (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Goat anti murine tyrosinase igg antibody, by Thermo Fisher Scientific (2 mentions)
Tc191 500mg, by HiMedia (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Dcfh da, by Merck Group (1 mentions)
Dmem f12, by HiMedia (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ro 32 0432, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Pd98059, by Merck Group (1 mentions)
Phosphate buffered saline rm7385 1pk, by HiMedia (1 mentions)
Potassium dichromate, by Merck Group (1 mentions)
Lead nitrate, by Merck Group (1 mentions)
Pb no3 2, by Merck Group (1 mentions)
Dimethyl sulphoxide dmso, by HiMedia (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
K2cr2o7, by Merck Group (1 mentions)
5 protocols using trypsin edta solution 1x
1
Cell Viability and Oxidative Stress Assays
2
Cultivation of Mouse Leydig Cells
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Mouse Leydig (TM-3) cells obtained from the American Type Culture Collection (ATCC®, USA) were cultured using a standard sterile cell culture technique. Medium consisted of Dulbecco’s Modified Eagle’s Medium F12 (DMEM/F12, 1:1 Mixture, Himedia) with 10% Fetal Bovine Serum (FBS; Gibco TM) and 1% Penicillin-Streptomycin (Sigma). The cells were maintained at 37°C in humidified air containing 5% CO2. Routinely, TM-3 cells were cultured in T-75 culture flasks containing 15–18 ml complete cell culture medium. They were sub-cultured, when 80–90% confluent, cells were harvested by using 0.25% Trypsin-EDTA 1X Solution (Himedia) and were used to conduct experiments.
3
Melanogenesis Regulation in Cell Culture
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For the present study, the compound, mushroom tyrosinase (lyophilized powder ≥ 1000 units/mg solid), and protein kinase inhibitors, protein kinase A (PKA) inhibitor (H89), protein kinase B (PKB) inhibitor (LY294002), protein kinase C (PKC) inhibitor (Ro-32-0432), and MEK1 inhibitor (PD98059) were purchased from Sigma-Aldrich, St. Louis, Missouri, United States. Goat anti-murine tyrosinase IgG antibody and Alexa Fluor® 594 donkey anti-goat IgG (H+L) (2 mg/mL) were purchased from Life Technologies, North America, United States. Dulbecco's Modified Eagle Medium (AT006A-5L), fetal bovine serum (RM10432-100 mL), Antibiotic Antimycotic Solution 100x (A002-20 mL), Trypsin-EDTA Solution 1x (TCL042-5 × 100 mL), MTT {[3-(4,5-dimethylthiazol-2-yl)]-2,5-diphenyltetrazolium bromide} (TC191-500MG), 4′,6-diamidino-2-phenylindole (DAPI) (TC229-5MG), phosphate buffered saline (RM7385-1PK), and Trypan blue, Certified (RM263-5G), were purchased from HiMedia Laboratories Pvt. Ltd., Mumbai.
4
Mushroom Tyrosinase Enzymatic Assay
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For the present study, the compound mushroom tyrosinase (lyophilized powder ≥1000 unit/mg solid), was purchased from Sigma-aldrich St. Louis, Missouri, United States. Goat anti-murine tyrosinase IgG antibody and IgG Alexa Flour ® 594 donkey anti-goat IgG (H + L) (2 mg/mL) was purchased from life technologies North America, United States. Dulbecco's Modified Eagle Medium (AT006A-5L) Fetal bovine serum (RM10432-100ML), Anitibiotic Antimycotic Solution 100X (A002-20ML), Trypsin-EDTA solution1X (TCL042-5 × 100ML), MTT {[3-(4, 5 Dimethylthiazol -2-y l)]-2, 5-diphenyltetrazolium bromide}(TC191-500MG), 4’, 6-diamidino-2-phenylindole (DAPI) (TC229-5MG), Phosphate buffered saline (RM7385-1PK) and Trypan blue, Certified (RM263-5G) were purchased from HiMedia Laboratories Pvt.Ltd. Mumbai.
5
Synthesis and Biocompatibility of Metal Nanoparticles
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All the chemicals used were either AR or GR grades. Lead nitrate, Pb (NO3)2 (99%), potassium dichromate, K2Cr2O7 (99%), ethanol (99%), hydrochloric acid, HCl and sodium hydroxide, NaOH (98%) were obtained from Merck, India. Ferric chloride hexahydrate, FeCl3 . 6H2O (97%) and ferrous chloride tetra hydrate, FeCl2 . 4H2O (97%) were purchased from CDH, India. L-Cysteine (L-Cyst 98%) was purchased from Sigma Aldrich. Chemicals including Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), antibiotic-antimycotic solution 100X, Trypsin-EDTA Solution 1X, dimethyl sulphoxide (DMSO), Dulbecco’s phosphate buffered saline (DPBS) 1X and 3-(4, 5-dimethyle-2-thiazolyl)-2 and 5-diphenyl-2H-tetrazolium bromide (MTT) for biocompatibility studies were procured from HiMedia Laboratories Pvt. Ltd. Tissue culture treated plate (96-wells) and T-20 flask received from Costar 3599 and Nucleon respectively. MTT stock solution (5 mg/mL) was prepared in DPBS by dissolving MTT with the help of vortex. All chemicals were used as received without further purification.
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