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2 protocols using neutral red dye

1

Melanogenesis Inhibition Protocol

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4‐HR (Kumar Organics, Bangalore, India), 4‐butylresorcinol (4‐BR, TCI, Chennai, India), 4‐phenylethylresorcinol (4‐PER, Spechem, Chennai, India), hydrogen phosphate and potassium dihydrogen phosphate (S.D Fine‐Chemical Ltd, Mumbai, India), hydroquinone and potassium chloride (Merck, Darmstadt, Germany), sodium chloride (Fischer Scientific, Waltham, USA), ethanol absolute 99.9% (Commercial alcohols, Ontario, Canada) and tissue solubiliser ‐Solvable (Perkin Elmer, Boston, USA). Kojic acid, arbutin, niacinamide, disodium triton X 100, RIPA buffer, 3‐methyl benzothiazolinone‐2(3H)‐hydrazone (MBTH), 3,4‐dihydroxyphenylalanine (DOPA), tyrosine, phenylmethanesulphonyl fluoride (PMSF), glacial acetic acid, calcein‐AM, and dimethyl sulphoxide (DMSO) were procured from Sigma‐Aldrich, St. Louis, USA, and neutral red dye from HiMedia, Kennett Square, USA. Sf9 insect cell line, High Five (H5) insect cell line, and Cellfectin II were purchased from Invitrogen, Waltham, USA; BacPAK baculovirus rapid titer kit (Clontech, Mountain View, USA); procollagen Type I C‐peptide (PIP) EIA kit (Takara, San Jose, USA); micro‐BCA protein assay kit (Thermo Scientific, Waltham, USA).
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2

Cell Culture Protocol with Reagents

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Cell culture plates and other plasticwares used in the current study were obtained from Tarsons Product Pvt. Ltd. Sigma-Aldrich, USA. The fetal bovine serum (FBS), trypsin-EDTA (0.25%), and Dulbecco’s Modified Eagle Medium (DMEM) were obtained from Sigma-Aldrich in the United States. Neutral red dye, MTT, and other chemicals and solvents were of analytical grade and procured from Hi-Media, USA.
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