Once confluent, cell monolayers were washed twice with warm phosphate-buffered saline (PBS) buffer (Sigma-Aldrich) to remove antibiotics. Then, CLAB cells were cultured in a 12-well cell culture plate (Corning Incorporated) at a concentration of 4 × 105 cells/well in DMEM medium containing P/S and L-glutamine (1.5 mL) in a controlled humidified atmosphere with 5% CO2 at 37% for 48 h. The probiotic strain L. reuteri B1/1 in MRS broth (Merck KGaA) was centrifuged at 2400 RPM for 10 min and resuspended in DMEM advanced media supplemented with 2 mM L-glutamine (both from Life Technologies) and without antibiotics. A 1 mL volume of the probiotic bacterial suspension at a concentration of 107 (LR7) and 109 (LR9) CFU/mL was added to the CLAB cells. As a control (C), the same medium but without bacteria was used for cell exposure. Plates were incubated for 2 h and 4 h under a controlled atmosphere (5% CO2 at 37%). All treatments and controls were performed in three independent replicates. After incubation, the cells were immediately washed with ice-cold sterile PBS buffer (Sigma-Aldrich).
Phosphate buffered saline buffer
Manufactured by Merck Group
Sourced in United States
Phosphate-buffered saline (PBS) is a commonly used buffer solution in biological research and laboratory applications. It is a balanced salt solution containing sodium phosphate and sodium chloride, with a pH of 7.4. The primary function of PBS is to maintain a physiologically relevant environment for various biological samples and experiments, such as cell culture, immunoassays, and protein purification.
Automatically generated - may contain errors
Lab products found in correlation
Formic acid, by Thermo Fisher Scientific (3 mentions)
12 well cell culture plate, by Corning (1 mentions)
Mrs broth, by Merck Group (1 mentions)
Pbs buffer, by Merck Group (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Merck Group (1 mentions)
Dextran sulfate, by Merck Group (1 mentions)
Milliq integral, by Merck Group (1 mentions)
Ezcount mtt cell assay kit, by HiMedia (1 mentions)
His tag antibody, by Thermo Fisher Scientific (1 mentions)
Sds page protein loading buffer, by Thermo Fisher Scientific (1 mentions)
Tris edta buffer, by Merck Group (1 mentions)
Proteinase k, by Merck Group (1 mentions)
Lysophosphatidic acid, by Enzo Life Sciences (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
11 mercaptoundecanoic acid, by Merck Group (1 mentions)
Milli q, by Merck Group (1 mentions)
Microcon ym 30, by Merck Group (1 mentions)
Hplc grade acetonitrile, by Thermo Fisher Scientific (1 mentions)
18 protocols using phosphate buffered saline buffer
1
Probiotic Lactobacillus reuteri Effects on Colon Cells
2
Lipid and Polymer Preparation for Biophysical Experiments
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DOPG was purchased from Avanti Polar Lipids and delivered as sodium salt solutions in chloroform. Dextran sulfate (as a powder, from Leuconostoc mesenteroides, >500 kDa) was purchased from Sigma-Aldrich. Milli-Q water was produced using a Milli-Q Integral water purification system (Merck Millipore; the electrical resistivity of the water was measured and was always above 18 MΩ.cm). Phosphate buffered saline (PBS) buffer was supplied by Sigma-Aldrich as tablets yielding 10 mM phosphate buffer containing 2.7 mM potassium chloride and 137 mM sodium chloride.
3
Fabrication and Evaluation of Diclofenac-loaded MOF Hydrogel
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The polydimethylsiloxane (PDMS) (SYLGARD 184 silicone elastomer and curing agent) used for device fabrication was purchased from Dow Corning Pvt. Ltd., Bangalore, Karnataka, India. The primary metal salt, Cobalt (II) Nitrate hexahydrate (Co(NO3)2.6H2O), and linker, 2-Methylimidazole (2-MIM), were purchased from AVRA Pvt. Ltd., Bangalore, Karnataka, India. The secondary metal salt, Zinc Nitrate hexahydrate (Zn(NO3)2.6H2O), metal salt (linker), Calcium chloride (CaCl2), and biopolymer, sodium alginate, were purchased from NICE Chemicals Pvt. Ltd., Bangalore, Karnataka, India. The drug diclofenac sodium was purchased as tablets from CIPLA, and the tablets were crushed into a powdered form. The MTT assay kit (EZcount MTT Cell Assay Kit) was purchased from HiMedia Laboratories Pvt. Ltd., Bangalore, Karnataka, India. Dulbecco’s Modified Eagle Medium (DMEM), Dimethyl Sulfoxide (DMSO), and phosphate-buffered saline (PBS) buffer were purchased from Sigma Aldrich (India) Pvt. Ltd., Bangalore, Karnataka, India. MCF-7 breast cancer cells were acquired from the Department of Biotechnology, JAIN (Deemed-to-be-University), Karnataka, India.
4
Bacterial Cell Wall Protein Binding Analysis
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A 20 ml of E. coli grown in LB medium, Clostridium thermocellum and C. cellulolyticum grown on 5 g/L cellobiose in VM medium were collected and centrifuged respectively. For each strain, the cell pellets were treated with NaN3/Ca2+ and washed three times with 50 mM Phosphate-buffered saline (PBS) buffer (Sigma-Aldrich, St. Louis, MO, United States). Then, 50 μg purified X2-N, ΔX2-N, and CBM3a proteins were incubated with collected cell pellets respectively in the PBS buffer at 4°C for 12 h. After centrifugation, the pellet from each incubation was resuspended in the SDS-PAGE Protein Loading Buffer (Thermo Fisher, Waltham, MA, United States) and boiled for 10 min. Finally, the 6x-His Tag antibody (R930-25, Thermo Fisher, Waltham, MA, United States) was used to detect the binding between proteins and cell wall by Western blotting.
5
Colorimetric Cytotoxicity Assay Protocol
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PDG (Sigma-Aldrich, CAS 82-89-3) was dissolved in ethanol and the 10 mg/mL primary stock solution was prepared and stored at -20 °C. For all the tests, the PDG stock solution was used to prepare 5 to 100 μM concentrations. 3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) powder, Phosphate-buffered saline (PBS) buffer, Tris-ethylenediaminetetraacetic acid (Tris-EDTA) buffer, and proteinase K were bought from Sigma-Aldrich Company and all the other chemicals and solutions were bought from Merck (Germany).
6
Quartz Crystal Biosensor Fabrication
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Sodium dodecyl sulfate, phosphate-buffered saline (PBS) buffer, and goat serum were all purchased from Sigma-Aldrich, Oakville, ON, Canada. Lysophosphatidic acid was obtained from Enzo Life Sciences via FroggaBio Inc., ON, Canada and Nitrogen as a pressurized gas from Praxair, ON, Canada. Quartz crystals (AT-cut, 13.5 mm in diameter, 9 MHz fundamental frequency) with gold electrodes in place were purchased from Lap-Tech Inc. Bowmanville, ON, Canada. The devices were systematically handled with thoroughly pre-cleaned stainless steel tweezers in order to minimize any external contamination. The synthesis of 2-(2-mercaptoethoxy)-ethan-1-ol (HS-MEG-OH) has been described elsewhere [20 (link)].
7
Multimodal CSFV E2 Protein Detection
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CSFV E2 glycoprotein, rabbit anti-CSFV E2 antibody, AP-conjugated goat anti-rabbit IgG and BCIP/NBT substrate were obtained from Beijing Bo Sheng Bio-technology Co., Ltd. Anti-PRV (porcine pseudorabies virus) antibody, anti-PCV2 (porcine circovirus type2) antibody and fluorescein isothiocyanate (FITC)-labeled anti-CSFV E2 antibody were acquired from Green biological technology corporation, Ltd., China. 11-Mercaptoundecanoic acid, 1-ethyl-3-(3-dimethylaminopropyl) carbodimide hydrochloride (EDC), N-hydroxysulfosuccinimide (NHS), bovine serum albumin (BSA, 99%) and phosphate buffered saline (PBS buffer, pH = 7.4) were purchased from Sigma-Aldrich Corporation (Saint Louis, MO, USA).
8
Extraction and Identification of Poria cocos Compounds
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Poria cocos was purchased from Sichuan Province (China) and was identified by Professor Chunming Liu (Changchun Normal University, Changchun, China). AChE was obtained from Sigma (St. Louis, MO, USA). We also used a Microcon YM−30 (Millipore, Billerica, MA, USA) ultrafiltration chamber with the molecular weight cutoff of 30,000 Da. The phosphate−buffered saline (PBS) buffer was purchased from Sigma (St Louis, MO, USA). HPLC−grade acetonitrile and formic acid were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Acetylthiocholine Chloride was purchased from Shanghai Yuanye Biotechnology Co., LTD. (CAS: 6050-81-3, Shanghai, China). Huperzine A was purchased from Shanghai Yuanye Biotechnology Co., LTD. (CAS: 102518−79−6, Shanghai, China). Chollne semiconductor solution was purchased from Shanghai Aladdin Biochemical Technology Co., LTD. (CAS: 123−44−1, Shanghai, China). The solvents and all other chemicals used in the study were of analytical grade and were purchased from Beijing Chemical Engineering Company (Beijing, China). Water was purified using a Milli−Q (Millipore, Boston, MA, USA) water purification system.
9
Quercetin Encapsulation in Polymer Nanoparticles
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Iron (III) chloride hexahydrate (97%) was purchased from Alfa Easar (Ottawa, ON, Canada). Ammonium acetate and polyEthylene glycol (PEG, Mw = 4000 Da) were obtained from Sigma Aldrich (St. Louis, MO, USA). Ethylene glycol and ethanol (96%) were purchased from Lach-ner (Neratovice, Czech Republic). Compressed nitrogen was purchased from Messer (Bad Soden am Taunos, Germany). Silicon oil was received from Acros organics (Waltham, MA, USA). Phosphate buffered saline (PBS) buffer (PBS tablets, pH 7.4, Ic = 150 × 10−3 mol dm−3) were purchased from Sigma Aldrich (St. Louis, MO, USA). Deionized water Millipore mili Q-H2O was used to prepare the PBS medium. Quercetin (≥99%) was supplied by Lach-ner (Neratovice, Czech Republic). A molecular weight cut-off dialysis bag (MWCO, 8Kd) was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
10
Measuring Expression of HFQ Variants
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To confirm the expression level of hfq variants, the gfp gene was cloned downstream of the hfq coding sequence as a fusion protein. The fluorescence intensity of GFP was measured by Guava Easycyte HT BG Flow Cytometer (Millipore, Billerica, MA, USA) to determine the expression level of hfq variants. For fluorescence measurement, harvest cells were washed and resuspended in 1× phosphate-buffered saline (PBS) buffer at pH 7.4 (Sigma-Aldrich, St. Louis, USA), and subsequently diluted 1000 times with 1× PBS buffer. All experiments were performed in triplicate.
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