Microarray kit (PAHS_143Z), RT2 first strand kit and RNA later™ were purchased from Qiagen (USA). Minimum Essential Medium (MEM), fetal bovine serum, 0.25% trypsin-EDTA, and 100X antibiotic and antimycotic mix were from HiMedia (India). Arecoline hydrobromide, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) kit, 2′,7′ –dichlorofluorescin diacetate (CM-H2DCFDA), sodium fluoride (NaF) and N-acetyl-L-cysteine (NAC) were purchased from Sigma-Aldrich (USA). Antibodies were purchased from Santa Cruz (USA) and Abcam (USA). The list of all the antibodies and primers used are provided in Supplementary Tables 1 and 2, respectively. TRIzol®, reverse transcription kit Superscript-RT and lipofectamine was purchased from Invitrogen (USA). KAPA SYBR FAST qPCR KIT Master Mix (2X) was procured from Kapa Biosystems (USA).
Trypsin edta
Manufactured by HiMedia
Sourced in India
Trypsin-EDTA is a cell culture reagent used to dissociate adherent cells from cell culture surfaces. It is a mixture of the proteolytic enzyme trypsin and the chelating agent EDTA, which work together to break down the extracellular matrix and cell-to-cell adhesions, allowing cells to be detached and harvested for further experimentation or subculturing.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by HiMedia (12 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by HiMedia (5 mentions)
Acridine orange, by Merck Group (3 mentions)
Propidium iodide, by Merck Group (3 mentions)
Antibiotic antimycotic solution, by HiMedia (3 mentions)
Rpmi 1640, by HiMedia (3 mentions)
Dulbecco s modified eagle s medium dmem, by HiMedia (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Dmem media, by HiMedia (2 mentions)
Formaldehyde, by Merck Group (2 mentions)
Phosphate buffer saline (pbs), by HiMedia (2 mentions)
Curcumin, by Merck Group (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by HiMedia (2 mentions)
Dulbecco s modified eagle s medium, by Merck Group (2 mentions)
Methanol, by Merck Group (2 mentions)
Ethyl acetate, by Merck Group (2 mentions)
Potato dextrose agar (pda), by HiMedia (2 mentions)
Resazurin, by Merck Group (2 mentions)
37 protocols using trypsin edta
1
Arecoline-induced Oxidative Stress Assay
2
Proliferation Kinetics of PDLSCs
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Cells at passage 3 were placed at 500 cells/cm 2 in a 12-well culture plate (Thermo Fisher Scientific) and cultured for 12 days in a basal culture medium. The cell culture medium was replenished every 3 days. PDLSCs in every three wells were washed with DPBS and detached using 0.25% trypsin-EDTA (HiMedia) on days 3, 6, 9, and 12, and counted using a hemocytometer. The experiment was performed in triplicates and the proliferation rate was calculated.
3
Adipogenic Differentiation Assay Protocol
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Syringic acid (SA, Sigma), Dulbecco's Modified Eagle's Medium (DMEM, HiMedia), phenol red-free Dulbecco's modified eagle's medium (HiMedia), insulin (HiMedia), dexamethasone (DEX, HiMedia), 0.25% trypsin-EDTA (HiMedia), Dulbecco's Phosphate Buffered Saline (DPBS, HiMedia), 3-isobutyl-1-methylxanthine (IBMX, Sigma), dimethyl sulphoxide (DMSO), free glycerol reagent (Sigma), foetal bovine serum (FBS, Gibco®), 100X antibiotic-antimycotic (Gibco®), thiazolyl blue tetrazolium bromide (MTT, HiMedia), β-nicotinamide adenine dinucleotide (reduced) disodium salt (β-NADH, SRL Pvt. Ltd), Oil-Red-O (ORO, HiMedia), nitroblue tetrazolium chloride (NBT, HiMedia), fatty acid-free bovine serum albumin (BSA, HiMedia) . All reagents and chemicals used were of tissue culture grade.
4
Synthesis and Characterization of Novel Complexes
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In this work,
commercial-quality reagents and solvents were used. All of the chemicals
and biochemicals were procured from Sigma-Aldrich Chemical Ltd, Merck.
All cell lines were purchased from NCCS, Pune. DMEM medium, 1% penicillin,
streptomycin, and 1% Glutmax were bought from Gibco. 10% Fetal bovine
serum and 0.25% trypsin-EDTA were obtained from Himedia and Thermo
Fisher Scientific, respectively. NMR spectra were recorded on a 400
MHz Advance Bruker DPX spectrometer with tetramethylsilane (TMS) as
the internal standard. An Elchem Microprocessor-based DT apparatus
was used to measure the melting points of the complexes. Infrared
(IR) spectra were recorded on a Shimadzu Affinity FT-IR spectrometer
in the range of 4000–400 cm–1. The mass spectra
of the synthesized compounds were recorded on Applied Biosystems (API-4000
ESI-mode), using methanol as the solvent. UV–visible and fluorescence
spectra were recorded on a JASCO V-760 spectrometer and Hitachi F7000
fluorescence spectrophotometer, respectively. A TDS conductometer
was used to measure the conductivity. An Elisa reader and 96-well
plates were used for the MTT assay.
commercial-quality reagents and solvents were used. All of the chemicals
and biochemicals were procured from Sigma-Aldrich Chemical Ltd, Merck.
All cell lines were purchased from NCCS, Pune. DMEM medium, 1% penicillin,
streptomycin, and 1% Glutmax were bought from Gibco. 10% Fetal bovine
serum and 0.25% trypsin-EDTA were obtained from Himedia and Thermo
Fisher Scientific, respectively. NMR spectra were recorded on a 400
MHz Advance Bruker DPX spectrometer with tetramethylsilane (TMS) as
the internal standard. An Elchem Microprocessor-based DT apparatus
was used to measure the melting points of the complexes. Infrared
(IR) spectra were recorded on a Shimadzu Affinity FT-IR spectrometer
in the range of 4000–400 cm–1. The mass spectra
of the synthesized compounds were recorded on Applied Biosystems (API-4000
ESI-mode), using methanol as the solvent. UV–visible and fluorescence
spectra were recorded on a JASCO V-760 spectrometer and Hitachi F7000
fluorescence spectrophotometer, respectively. A TDS conductometer
was used to measure the conductivity. An Elisa reader and 96-well
plates were used for the MTT assay.
5
Cytotoxicity Evaluation of Casein Protein
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Casein protein was purchased from Sigma Aldrich (C3400). Acetone was purchased from Merck (1.00983.0511). MTT dye (TC191), glutaraldehyde 25% w/v solution (RM5927), propidium iodide (PI, TC252), streptomycin penicillin solution (A018), Dulbecco modified Eagle medium (DMEM, GIBCO, AL007S), fetal bovine serum (FBS, RM9955) and trypsin–EDTA were purchased from Himedia, as well as dialysis membrane 12 kD (D6191). The annexin V-PE/7-AAD kit (BD-Bioscience) and JC-1 cyanine dye (Abcam) were used for apoptosis and mitochondrial membrane potential analysis via fluorescence-activated cell sorting (FACS) respectively. DAPI (Himedia) was used for nuclear staining and 2′7′-dichlorofluorescein diacetate (Himedia) was used as a fluorescent dye for ROS detection respectively.
6
Assessing MRSA Infection and Probiotic Treatment
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Caco-2 cell monolayer was washed twice with 1X PBS and the cells were suspended in DMEM. The Caco-2 cell monolayer was infected with MRSA (MOI 1:10) and incubated at 37°C for 2 h with 5% CO2–95% air. After 2 h, the cells monolayer was washed and treated with L. fermentum (MOI 1:100) for 1 h at 37°C. After treatment; the cells monolayer was washed and incubated for 24 h at 37°C with 5% CO2–95% air. After incubation, the Caco-2 cells were trypsinized using Trypsin- EDTA (HiMedia) for 3 min and inactivated by adding 20% FBS. The trypsinized cells were centrifuged and suspended in 1X PBS. PI (5 mg/ml) was added to each sample, and cell viability was analyzed using FACSAria III.
7
Assessing MRSA-Infected Caco-2 Cell Viability
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Live and dead cell assay was performed to determine the viability of MRSA infected Caco-2 cells. Caco-2 cell monolayer was washed twice with 1X PBS, and the cells were suspended in DMEM medium. The cells were infected with MRSA at an MOI of 1:10 and further incubated for various time points (4, 16, and 24 h) at 37°C. At each time point, the cells were trypsinized using Trypsin-EDTA (HiMedia) for 3 min followed by addition of 20% FBS for inactivation. The trypsinized cells were centrifuged and suspended in 1X PBS. Propidium iodide (PI) (5 mg/ml) was added to each sample, and live/dead cells were quantified using FACSAria III. Uninfected cells were used as a control.
8
Preparing Folate-Conjugated Doxorubicin
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cholesterol chloroformate, folic acid, and cholesterol were purchased from Sigma Aldrich (Mumbai, India); DOX was a gift from Naprod Life Sciences Ltd. (Mumbai, India); dimethyl sulfoxide (DMSO), dicyclohexyl carbodiimide (DCC) n-hydroxy succinamide (NHS), diethyl ether, hexane, ethyl acetate, butanol, glacial acetic acid, and sodium dodecyl sulfate solution (SDS) were purchased from S.D. Fine Chem Ltd. (Mumbai, India). Trehalose (Hayashibara, Japan) was supplied as gift sample by Gangwal Chemicals Pvt. Ltd (Mumbai, India). Folate-free DMEM medium, trypsin EDTA, and fetal bovine serum (FBS) were obtained from Himedia Ltd. (Mumbai, India). All other chemicals and reagents used were of analytical grade. All the animal studies were approved by independent ethics committee of Bhabha Atomic Research Centre, Mumbai, India. All the procedures involved in the animal studies were performed in accordance with the recommendations of NIH guidelines for the proper use and care of laboratory animals.
9
Biochemical Assays for Cell Viability
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Dulbecco’s modified eagle medium (DMEM), Roswell Park Memorial Institute 1640 (RPMI-1640), penicillin, streptomycin, and trypsin-EDTA (ethylenediaminetetraacetic acid) were procured from Himedia (Mumbai, Maharashtra, India). Crystal violet, dimethyl sulfoxide (DMSO), and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) were obtained from SRL Diagnostics (Mumbai, Maharashtra, India). The 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), 4′,6-diamidino-2-phenylindole (DAPI), 5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethyl benzimidazolyl carbocyanineiodide (JC-1), acridine orange, agarose, alexa fluor 488, ethidium bromide, fetal bovine serum (FBS), and propidium iodide were obtained from Invitrogen (Carlsbad, CA, USA). Bcl-xL antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) while GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was obtained from Abgenex (Bhubaneswar, Odisha, India).
10
Synthesis and Characterization of Fluorescent Polymeric Nanoparticles
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Sodium hydride, 1-dodecanethiol, 4,4-azobis(4-cyanovaleric acid), deuterated chloroform (CDCl3), 2-hydroxy ethyl methacrylate (HEMA), polyethylene glycol methyl ether methacrylate −300 (PEGMA), pyrene, and MTT were purchased from Sigma Aldrich (Germany) and were used without further purification. Stearic acid, DMEM media, and molecular biology grade water, fetal bovine serum (Brazil origin, EU approved), and trypsin-EDTA were purchased from Himedia laboratories private limited (Mumbai, India). Doxorubicin HCl was purchased from Tokyo Chemical Industry, (Japan). Dicyclohexylcarbodimide (DCC), 4-dimethylaminopyridine (DMAP) were purchased from SRL chemicals (Mumbai, India). 2,20-Azo-bis-(isobutyronitrile) (AIBN) were purchased from SRL and were recrystallized from methanol before use. Fluorescent probes pyrene and DAPI were purchased from Sigma Aldrich (Germany). All the solvents were used as purchased without any further purification.
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