Dubos medium was purchased from Difco, Detroit, MI, USA. Luria–Bertani (LB) broth, Dulbecco’s Modified Eagle’s Medium (DMEM), Roswell Park Memorial Institute (RPMI) 1640 medium, fetal bovine serum (FBS), minimum essential medium (MEM), sodium pyruvate, sodium nitrate, and amino acids were procured from HiMedia, Mumbai, India. Ampicillin, gentamicin, rifampicin, dimethyl sulfoxide (DMSO), 4′,6-diamidino-2-phenylindole (DAPI), phorbol myristate acetate, and MTT dye (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) were obtained from Sigma-Aldrich (St Louis, MO, USA).
Amino acids
Manufactured by HiMedia
Sourced in India
Amino acids are the fundamental building blocks of proteins. They are organic compounds consisting of a central carbon atom, an amino group, a carboxyl group, and a side chain. Amino acids play a crucial role in various biological processes and are essential for the proper functioning of the human body.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 media, by HiMedia (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Apoptosis detection kit, by Merck Group (2 mentions)
Coumaric acid, by Merck Group (2 mentions)
Fluorescein isothiocyanate fitc conjugated annexin 5, by Merck Group (2 mentions)
Rnase a, by Merck Group (2 mentions)
Luminol, by Merck Group (2 mentions)
Bradford reagent, by Merck Group (2 mentions)
Methylthiazolyldiphenyl tetrazolium bromide mtt, by Sisco Research Laboratories (2 mentions)
Dmem media, by HiMedia (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
1640 medium, by HiMedia (1 mentions)
Sodium pyruvate, by HiMedia (1 mentions)
Sodium nitrate, by HiMedia (1 mentions)
Mtt dye, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Gentamicin, by Merck Group (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by HiMedia (1 mentions)
Luria bertani broth, by HiMedia (1 mentions)
5 protocols using amino acids
1
Cell Culture Media and Reagents
2
Cell Viability and Apoptosis Assay
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RPMI-1640 media, DMEM media and other necessary chemicals like antibiotics, amino acids, etc. were purchased from HIMEDIA (Mumbai, India). Fetal bovine serum (FBS) was purchased from HyClone (Thermo Scientific Hy-Clone, Logan, Utah, USA). Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was purchased from Sisco Research Laboratory (Mumbai, India). Cisplatin, ribonuclease, Fluorescein isothiocyanate (FITC) conjugated Annexin V, apoptosis detection kit, Z-vad FMK, RNaseA, Bradford reagent, luminol and coumaric acid were purchased from Sigma (Missouri, USA). Antibodies were purchased from abcam (Cambridge, UK) and Cell Signaling Technology (Danvers, MA, USA). Other essential chemicals used in this study were of the analytical grade.
3
Synthesis and Characterization of Curcumin-Functionalized Nanoparticles
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Magnesium(II) acetate tetrahydrate (99%), zinc(II) acetate dihydrate (99.5%), N-hydroxyl succinimide (97%), 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (EDC∙HCl) extra pure (99%), 3-carboxybenzeneboronic acid (97%), potassium hydroxide pellets, disodium dihydrogen phosphate dihydrate, potassium dihydrogen phosphate, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and sodium chloride were purchased from Sisco Research Laboratory (Mumbai, India). 3-aminopropyl triethoxysilane (APTES) with a purity of 99% was purchased from Sigma-Aldrich (St. Louis, Missouri, USA). Ethyl alcohol (EtOH) and dimethyl sulfoxide (DMSO) were purchased from Merck (Kenilworth, New Jersey, United States). Curcumin, DMEM media, RPMI-1640 media, amino acids and antibiotics were purchased from Hi-Media (Mumbai, India). Fetal bovine serum (FBS) was purchased from Thermo Scientific Hy-Clone (Logan, Utah, USA).
4
Yeast Strain Construction and Culture Conditions
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The S. cerevisiae strains used in this study were MLC30M (MATa trp1-1 leu2-3,112 his3-11,15 ura3-1 ade2-1 can1-100 yrs1::HIS3 yrr1::TRP1 pdr1::hisG pdr3::hisG) (NBRP, Japan)29 (link),30 and BY4741 (MATa leu2Δ0 ura3Δ0 his3-Δ1 met15Δ0) (NBRP, Japan)31 ,32 . The gene deletion mutants of BY4741 were purchased from Euroscarf, Germany. The deleted genes were replaced by kanMX4. Yeast cells were cultured at 30 °C in Yeast Peptone Dextrose (YPD) medium (10 g/L yeast extract, 20 g/L peptone, and 20 g/L dextrose) (HIMEDIA, India) for general growth and CLS screen of the mutants. Synthetic complete (SC) medium consisting of 2% glucose (HIMEDIA, India), 0.67% yeast nitrogen base with ammonium sulphate (HIMEDIA, India), and amino acids (HIMEDIA, India) at appropriate concentrations and lacked certain amino acids (uracil, leucine) were used for plasmid selection and protein expression assays. The concentrations of amino acids in SC medium are listed in Table 1 . Yeast transformation was performed as described33 (link), and transformants were selected from synthetic complete minus uracil (SC-Ura) agar.
Final concentrations of amino acids in SC medium.
| Amino acid | Final concentration in SC medium (mg/L) |
|---|---|
| L-Isoleucine | 30 |
| L-Valine | 150 |
| Adenine sulphate | 20 |
| L-Arginine | 20 |
| L-Lysine | 30 |
| L-Methionine | 20 |
| L-Phenylalanine | 50 |
| L-Threonine | 200 |
| L-Tyrosine | 30 |
| L-Histidine | 24 |
| L-Tryptophan | 24 |
| L-Leucine | 36 |
5
Oxidative Stress-Induced Apoptosis Pathways
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RPMI-1640 media and other necessary chemicals like antibiotics, amino acids etc were purchased from HIMEDIA (Mumbai, India) and fetal bovine serum (FBS) from HyClone (Thermo Scientific Hy-Clone, Logan, Utah), respectively. tBHP was purchased from Sigma-Aldrich and Methylthiazolyldiphenyl-tetrazolium bromide (MTT) were purchased from Sisco Research Laboratory (Mumbai, India). Fluorescein isothiocyanate (FITC) conjugated Annexin V, Apoptosis detection kit, RNaseA, Bradford reagent, Luminol, Coumaric acid and LY294002 (a PI3K inhibitor) were also purchased from Sigma (Missouri, USA). Antibodies such as anti Caspase-3 (ab47131), anti Caspase-8 (ab25901), anti Bid (ab77815), anti Bcl2 (ab7973), anti cytochrome c (ab76237), anti Nrf2 (ab31163), anti HO1 (ab13243), anti SOD2 (ab13533), Phospho JNK (ab4821), anti Bax (ab32503), anti PI3k (ab74136), anti Akt (ab17785), Phospho Akt (ab23509), HRP (ab97051) were purchased from Abcam (Cambridge, UK). Phospho BAD (#9291), anti mTOR (#2983), anti GSK 3β (5338), anti Cyclin D1 (#2978), anti PARP (#9532), anti Caspase-9 (#9508), anti Apaf-1 (#8723), anti Bax (#2772), Anti NFκB (#3034), anti β-Actin (#4970), anti Lamin B1 (#12586) was purchased from Cell Signaling Technology (Danvers, MA 01923). Other essential chemicals used in this study were of the analytical grade.
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