For studying apoptotic markers, protoplasts of C. auris MRL6057 were prepared as explained previously [21] (link). Briefly, cells grown for 8 h were then exposed for 4 h to ¼ MIC, ½ MIC and MIC of test compounds. For the purpose of washing and resuspending yeast cells different protoplast buffers (PB) were prepared. After exposure with test compounds, cells were washed and incubated in PB-1 (1 M sorbitol, (Sigma Aldrich Co., USA), 0.05 M tris base (Merck, Germany), 0.01 M MgCl2 (Sigma Aldrich Co., USA), 0.03 M DTT (Merck, Germany), pH 7.4) for 10 min at room temperature. Post-incubation, cells were harvested at 1500 rpm for 5 min and pellet was mixed and incubated in PB-2 (1 M sorbitol, 0.05 M tris base, 0.01 M MgCl2, 0.001 M DTT, pH 7.4) supplemented with lyticase enzyme (1 μg/mL; Sigma Aldrich Co., USA) at room temperature for 1 h. Next, cell suspensions were centrifuged, and pellets were resuspended and incubated in PB-3 (1 M sorbitol, 0.05 M tris base, 0.01 M MgCl2, pH 7.4) at room temperature for 20 min. Post-incubation cell suspension was centrifuged at 1500 rpm for 5 min and pellets with protoplasts were washed and mixed in fresh PBS and stored at 4 °C until further use.
Tris base
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Spain, China, India, Sao Tome and Principe, Italy, France
Tris base is a chemical compound used as a buffer in various laboratory applications. It is a white crystalline solid that is highly soluble in water. Tris base is commonly used to maintain a specific pH range in biological and biochemical experiments, such as in the preparation of buffers for gel electrophoresis, protein purification, and molecular biology techniques.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Merck Group (77 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (48 mentions)
Triton x 100, by Merck Group (44 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (44 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (43 mentions)
Acetic acid, by Merck Group (36 mentions)
Bovine serum albumin (bsa), by Merck Group (36 mentions)
Hydrochloric acid (hcl), by Merck Group (31 mentions)
Glycine, by Merck Group (30 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (29 mentions)
Tween 20, by Merck Group (28 mentions)
Dithiothreitol (dtt), by Merck Group (24 mentions)
Tris hcl, by Merck Group (22 mentions)
Trichloroacetic acid, by Merck Group (21 mentions)
Sodium hydroxide (naoh), by Merck Group (20 mentions)
Methanol, by Merck Group (17 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (17 mentions)
Urea, by Merck Group (16 mentions)
Potassium chloride (kcl), by Merck Group (16 mentions)
Mgcl2, by Merck Group (15 mentions)
352 protocols using tris base
1
Apoptotic Marker Analysis in C. auris
2
Isolation of Buchnera Endosymbionts from Pea Aphids
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Pea aphids (Acrythosiphon pisum strain LSR1) were grown as an all-female clone on Fava bean (Vicia faba) seedlings on 16h/8h light/dark cycles at 20°C. Once reaching adulthood, apterous adults were raised on Fava bean plants on 16h/8h light cycles and allowed to reproduce overnight. After seven days, all aphids, in the 4th larval instar and typically amounting to 5g, were removed from the Fava bean plants. Aphids were weighed and surface-sterilized in 0.5% NaClO solution, then rinsed twice in Ultrapure water (MilliporeSigma), each 30 s. Aphids were gently ground in a mortar and pestle in 40mL sterile Buffer A (25mM KCl (Sigma-Aldrich), 35mM Tris base (Sigma-Aldrich), 10mM MgCl2 (Sigma-Aldrich), 250mM anhydrous EDTA (Sigma-Aldrich), and 500mM Sucrose (Sigma-Aldrich) at pH 7.5). Aphid homogenate was vacuum filtered to 100μm, then centrifuged at 1500g for 10 min at 4°C. Supernatant was discarded, and the resulting pellet was resuspended in 20mL Buffer A and vacuum-filtered three times from 20μm, to 10μm, and finally to 5μm. The resulting filtrate was spun at 1500g for 30 min at 4°C and supernatant discarded. The resulting pellet was resuspended in 10mL Sucrose solution (300mM Sucrose (Sigma-Aldrich) and 100mM Tris base (Sigma-Aldrich)) then checked on a brightfield microscope for intact Buchnera cells. Buchnera cells remain intact while at 4°C for a maximum of 24h.
3
Isolation of Peripheral Blood Leukocytes
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The isolation of peripheral blood leukocytes (PBL) was done as previously described14 (link),17 (link) with methodology similar to one previously described30 . Whole blood was incubated in the same proportion with red blood lysis buffer solution [162.64 mM NH4Cl (Sigma-Aldrich), 9.98 mM Tris base (Merck, Darmstadt, Germany), pH = 7.2] for 10 min, with agitation, at room temperature. Cells were then passed through a 100-μm cell strainer, washed, and resuspended in complete RPMI medium: RPMI 1640 Medium supplemented with 10% foetal bovine serum (FBS) (FBS South America Premium, Ref. S181BH-500 from Biowest, Nuaillé, France), 85 units/mL penicillin, 85 μg/mL streptomycin, 62.5 ng/mL amphotericin B, 0.05 mM 2-mercaptoethanol and 10 mM HEPES (all from Sigma-Aldrich) after centrifugation at 300×g for 5 min.
4
Protein Gel Electrophoresis and Mass Spectrometry
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All reagents used were HPLC grade or electrophoresis grade. Acrylamide/bis-acrylamide 30% solution (37.5:1), ammonium bicarbonate (ambic), β-mercaptoethanol, Coomassie Brilliant Blue R250 (CBB), dithiothreitol (DTT), iodoacetamide (IAA), formic acid, glycerol 86–88%, sodium borohydride (NaBH4), sodium carbonate, sodium citrate tribasic dihydrate (HOC(COONa)(CH2COONa)2·2H2O), (N,N,N, N′-tetramethylethylenediamine (TMED), trifluoroacetic acid, tris-base, trypsin, and the Sigma Marker wide range 6.5–200 KDa were all from Merck (Barcelona, Spain). Acetonitrile, formaldehyde, methanol and sodium dodecyl sulfate (SDS) were supplied by Panreac Química SLU (Barcelona, Spain). Bromophenol-blue was purchased from Riedel-de Haen (Seelze, Germany). Pierce™ trypsin Protease, MS Grade was purchased from Thermo Fisher Scientific (Bremen, Spain). Hydrogen tetrachloroaurate (III) hydrate (HAuCl4·xH2O) (99.9% Au) (49% Au) at 10% w/v was acquired from Strem Chemicals (Kehl, Germany).
5
Chromatographic Analysis of Carbohydrates
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Each chemical and reagent used was of superior quality and according to the high-performance liquid chromatography (HPLC) standards. Sucrose, fructose, and maltose were purchased from the Indian based companies named Central Drug House (P) Ltd. New Delhi, India; Loba Chemie Pvt. Ltd. Mumbai, India; and Techno Pharmchem, New Delhi, India respectively. While, glucose was bought from th Chem-LabNV, Zedelgem, Belgium. However, 5-(hydroxymethyl) furfural, Mueller Hinton Broth, Sulforhodamine B sodium salt solution, and AgNO3 were purchased from the USA based company Sigma-Aldrich (St. Louis, MO, USA). HPLC standard grades water and acetonitrile were bought from Indian based company Chemie Pvt. Ltd. Mumbai, India. While, Roswell Park Memorial Institute (RPMI) 1640 medium and Penicillin-Streptomycin (10,000 U/mL ) were purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Fetal bovine serum (FBS), Trichloroacetic acid (TCA), acetic acid, and tris base were purchased from the Merck, Darmstadt, Germany.
6
Antimicrobial Evaluation of Selenium-Enriched Bacillus
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Meglumine antimoniate (MA, Glucantime) as control drug was prepared from Rhône, Poulenc, France. Penicillin and streptomycin were obtained from Alborz Pharmacy, Karaj, Iran and were stored at room temperature (25°C) until testing. MTT powder [3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide)], fetal calf serum (FCS) and RPMI-1640 medium with L-glutamine were purchased from Sigma-Aldrich, St Louis, MO. Selenium dioxide (SeO2), nutrient broth, nutrient agar, n-octyl alcohol, sodium dodecyl sulfate (SDS) and Tris base were purchased from Merck Chemicals (Germany). All other chemicals and solvents were of analytical grade. The microorganism used in this study was identified as Bacillus sp. MSh-1 by the methods described earlier (12 (link)). The organism was continuously maintained on nutrient agar plates complemented with 1.26 mM SeO2 using continuous sub-culturing every 14 days.
7
Immunohistochemical Analysis of MASP-2
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Kidney and liver samples were fixed in 4% paraformaldehyde (Merck) and embedded in paraffin. Blocks were cut in 1–4 µm sections, dewaxed, and rehydrated through a series of Tissue-Clear (Sakura ProHosp, Alphen an den Rijn, The Netherlands) and ethanol (99–70%) (Merck). Antigen retrieval was performed with TEG-buffer (10 mM TrisBase, 0.5 mM EGTA Tritiplex VI) (Merck) in a microwave oven for 20 min. Sections were washed in TBST (Merck) and blocked for 30 min in 3% Bovine Serum Albumin (BSA)/TBST (Merck), washed and blocked 10 min in hydrogen peroxide. Anti-MASP-2 (15-17-3) was applied undiluted/in culture supernatant overnight at 4 °C, washed, and incubated with HRP-conjugated goat-anti mouse-IgG (P0447; Dako) diluted 1:1000 for 1 h. Staining was visualized with 3.3′ diaminobenzidine (DAB)(Dako) and counterstained with Mayer's Hematoxylin (Merck). Images were taken with Olympus BX51 microscope with a DP26 camera and Cell Sens software (Olympus, Tokyo, Japan). Co-localization was investigated using anti-MASP-2, anti-AQP2 (collecting duct, principal cell marker, 1:200, 9882, Santa Cruz) and anti-ATP6V1B1 (collecting duct, intercalated cell marker, HPA031847, 1:50), respectively. AlexaFluor488 donkey-anti goat-IgG (Invitrogen) and AlexaFluor555 donkey-anti mouse-IgG (Invitrogen) were used as secondary antibodies.
8
Doxorubicin-loaded Hyaluronate Nanoparticles
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Sodium hyaluronate (HA, 10.8 kDa) was purchased from Lifecore Biomedical LLC (MN, USA). Oligonucleotides (ODN) (i-motif ODN (IMO, 5′-CCCTAACCCTAAAAAAA-NH2-3′) and i-motif binding ODN (IBO, 5′-TTTTTTTAGGGTTAGGG-NH2-3′)) were purchased from Bioneer (Daejeon, Korea). Doxorubicin (DOX) was purchased from MedChemExpress LLC (Monmouth Junction, NJ, USA). Polyethylenimine (1800 Da, PEI1.8k), agarose, 2-[4-(2-hydroxyethyl)piperazine-1-yl]ethanesulfonic acid (HEPES), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), ethidium bromide (EtBr), sodium acetate, 3-[4-5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium bromide (MTT), Stains-All, and N-hydroxysuccinimide (NHS) were purchased from Sigma Aldrich (St. Louis, MO, USA). Tris base, dimethyl sulfoxide (DMSO), and hydrochloric acid were purchased from Merck (Darmstadt, Germany). Tris-Ethylenediaminetetraacetic acid (TE) buffer was purchased from Promega (Madison, WI, USA). Dulbecco’s modified Eagle’s medium (DMEM), Dulbecco’s phosphate-buffered saline (DPBS), fetal bovine serum (FBS), penicillin/streptomycin (P/S), and trypsin-EDTA (0.25%) were purchased from Invitrogen (Carlsbad, CA, USA). All other chemicals were purchased and used without further purification.
9
Cytotoxicity Evaluation of Honey and Honey-AgNPs
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Additionally, 96-well plates were seeded with MCF-7, HeLa, and HePG2 cells, at approximately 105/well. Following 72 h exposure to honey and honey with AgNPs, the medium was changed to 150 L of 10% trichloroacetic acid (TCA) from Merck for 1 h at 4 °C; then, the cultures were washed five times with distilled water. Afterward, a 70 μL SRB solution (0.4% w/v) (Sigma Aldrich, St. Louis, MO, USA) was added for 10 min at room temperature in a dark location. The cells were washed three times with 1% acetic acid (Merck) and left overnight to air dry. The protein-bound SRB stain was dissolved by adding 150 μL of 10 mM Tris Base (Merck), and the O.D. was measured at 540 nm using a FluoStar Omega microplate reader (BMG Labtec, Ortenberg, Germany) [51 (link)].
10
Molecular Cloning Protocol Using BCL1 Enzyme
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Various materials, including Agarose (Cat No: 9012-36-6), Boric acid (Cat No: 10043-35-3), EDTA (Cat No: 60-00-4), and Tris- base (Cat No: 77-86-1), were prepared from Merck company (Germany). Also, Ladder 50 bp (Cat No: 32810), Self- Stain dye (Cat No: 25148), and PCR master mix (Cat No: 201289) were purchased from Sinaclone Co., and Extraction DNA Kit was obtained from Roje Co. The BCL1 restriction enzyme was supplied by New England Biolabs, Inc (Ipswich, MA, USA).
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