Phosphate buffered saline ph 7

Manufactured by Thermo Fisher Scientific

Sourced in United States, Switzerland

Phosphate-buffered saline (PBS) pH 7.4 is a commonly used buffer solution in biological and biochemical applications. It is an isotonic solution that maintains a physiologically relevant pH of 7.4. The solution contains sodium phosphate and sodium chloride, providing a stable environment for various experiments and processes involving cells, proteins, and other biological samples.

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37 protocols using phosphate buffered saline ph 7

HPLC Analysis of Mefloquine and Verapamil

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Mefloquine hydrochloride and verapamil hydrochloride (as the internal standard [IS]) were purchased from Sigma-Aldrich (Castle Hill, NSW, Australia). HPLC grade methanol, HPLC grade acetonitrile, phosphate buffered saline pH 7.4 (PBS) and triethylamine were purchased from Thermo Fisher Scientific (Macquarie Park, NSW, Australia).

Izes A.M., Kimble B., Norris J.M, & Govendir M. (2020). Assay validation and determination of in vitro binding of mefloquine to plasma proteins from clinically normal and FIP-affected cats. PLoS ONE, 15(8), e0236754.

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Cyclodextrin-based Formulations for LGK974

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Cyclodextrins (CD): α-cyclodextrin sulfobutyl ethers (αSBECD) and β-cyclodextrin sulfobutyl ethers (βSBECD) sodium salts were obtained from Ligand Pharmaceuticals, Inc. (San Diego, CA). CDs α-cyclodextrin (αCD), β-cyclodextrin (βCD) and (2-Hydroxypropyl)-β-cyclodextrin (HPβCD) were obtained from Sigma-Aldrich (Milwaukee, WI, USA). LGK974 was purchased from Selleckchem (Cambridge, MA, USA). Hydrochloric acid 37% ACS reagent, carboxymethylcellulose sodium salt (CMC), Tween-80, HPLC grade acetonitrile, trifluoroacetic acid 99% reagent plus, and dimethyl sulfoxide 99.9% ACS reagent were all obtained from Sigma-Aldrich (Milwaukee, WI, USA). Phosphate buffered saline, pH 7.4 was obtained from ThermoFisher Scientific (Cambridge, MA, USA). Deuterium Oxide (D₂O) was obtained from Cambridge Isotope Laboratories, Inc. (Cambridge, MA, USA).

Guimaraes P.P., Tan M., Tammela T., Wu K., Chung A., Oberli M., Wang K., Spektor R., Riley R.S., Viana C.T., Jacks T., Langer R, & Mitchell M.J. (2018). Potent in vivo lung cancer Wnt signaling inhibition via cyclodextrin-LGK974 inclusion complexes. Journal of controlled release : official journal of the Controlled Release Society, 290, 75-87.

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Formation of Fluorescent DOPC SUVs

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SUVs composed of 99% 18:1 DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) and 1% Atto633-DOPE (Atto633–1,2-dioleoyl-sn-glycero-3-phosphoethanolamin) were formed by mixing the lipids dissolved in CHCl₃ in a glass vial and subsequent solvent evaporation under a stream of nitrogen gas. The glass vial was then placed under a vacuum for 30 min to remove residual traces of solvent. Afterwards, the lipids were resuspended in phosphate buffered saline pH 7.4 (Thermo Fisher) at a final concentration of 1 mM lipids. The solution was vortexed for 10 min to trigger liposome formation. SUVs were then formed by extruding the liposome solution 13 times through a polycarbonate filter with a pore size of 50 nm (Avanti Polar Lipids). The SUVs were stored at 4 °C until use.

Zhan P., Jahnke K., Liu N, & Göpfrich K. (2022). Functional DNA-based cytoskeletons for synthetic cells. Nature Chemistry, 14(8), 958-963.

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Rat Pulmonary Microvascular Endothelial Cell Infection

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PMVECs from both male and female Fischer and male Sprague-Dawley rats were grown to 24 h post-confluence. The monolayer from an identically seeded counting plate was trypsinized and cells were counted with a Countess II Automated Cell Counter (Invitrogen, Carlsbad, CA, cat. no. AMQAX1000) according to manufacturer’s instructions. Bacteria from overnight Vogel-Bonner plates were suspended in 1x Phosphate Buffered Saline pH 7.4 (Invitrogen; Carlsbad, CA, cat. no. 10010–049) to an OD₅₄₀ of 0.25, previously determined to represent 2 × 10⁸ CFUs/mL, and diluted to an MOI of 20:1 in HBSS (Invitrogen; Carlsbad, CA, cat. no. 14025–134). Infected monolayers were incubated (37°C and 5% CO₂) for 4 h (PA103 and ΔPcrV), 5 h (PAO1, PA-808), or 7 h (ExoY⁺, ExoY^K81M). Supernatants were then collected, centrifuged at 4000 x g, and sterilized via passage through a PES 0.22 μm filter (EMD Millipore, Burlington, MA, cat. no. SLGP033RS). Vehicle control supernatant was generated as described for bacterial infection except bacteria were excluded from the PBS/HBSS solution used to treat the monolayers.

Voth S., Gwin M., Francis C.M., Balczon R., Frank D.W., Pittet J., Wagener B.M., Moser S.A., Alexeyev M., Housley N., Audia J.P., Piechocki S., Madera K., Simmons A., Crawford M, & Stevens T. (2020). Virulent Pseudomonas aeruginosa infection converts antimicrobial amyloids into cytotoxic prions. The FASEB Journal, 34(7), 9156-9179.

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Thioflavin T Amyloid Fibrillization Assay

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Thioflavin T (ThT) is a light-sensitive stain that is often used to study amyloid fibrilization kinetics or as an end-point read out to assess the quantity of fibrillar amyloid species in a given sample (50 (link)). A 25 mM ThT (Acros Organics; Geel, Belgium, cat. no. AC211760250) stock solution was prepared in 100% ethanol, vortexed thoroughly, and passed through a 0.22 μm syringe filter to remove clumps. Black clear-bottomed Corning Costar 96-well polystyrene plates (Corning Inc.; Corning, NY, cat. no. 3603) were used for assays. Supernatant samples were prepared as detailed for ‘Protein analysis’ and supernatants were diluted into 200 μL of 1x Phosphate Buffered Saline pH 7.4 (Invitrogen; Carlsbad, CA, cat. no. 10010–049) to a protein concentration of 5 mg/mL. ThT from the 25 mM stock solution was added to wells just prior to reading the plate at a final concentration of 0.49 μM. Fluorescence was measured on a Molecular Devices Spectramax iD5 Multi-Mode Microplate reader (425 excitation; 490 emission), 1 min orbital shaking prior to read, high gain, 400 ms integration time.

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Reagents and Material Acquisition

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Acetone, n-hexane, toluene, ethyl acetate, dichloromethane, methanol, acetonitrile, silica gel 60 GF254 plates, and silica gel 60 powder were purchased from Sigma-Aldrich Chemie (Schnelldorf, Germany). Mueller–Hinton broth (MHB), Lysogeny broth (LB), and agar were purchased from Carl Roth (Karlsruhe, Germany). Gentamicin sulfate was purchased from AppliChem (Darmstadt, Germany). Phosphate-buffered saline (PBS) pH 7.4 was purchased from Fischer Scientific (Schwerte, Germany) and Millipore water was prepared by the Milli-Q^® direct laboratory water purification system (Merk, Darmstadt, Germany).

Masota N.E., Ohlsen K., Schollmayer C., Meinel L, & Holzgrabe U. (2022). Isolation and Characterization of Galloylglucoses Effective against Multidrug-Resistant Strains of Escherichia coli and Klebsiella pneumoniae. Molecules, 27(15), 5045.

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Bioadhesive Hydrogel Tissue Bonding

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The starting materials included polyethylene glycol (M_w: 35 kDa), 2-hydroxy-1-[4-(2-hydroxyethoxy) phenyl]-2-methyl-1-propanone (Irgacure D-2959), lactide, triethylamine, acryloyl chloride, Tin(II) 2-ethylhexanoate [Sn(Oct)₂], dichloromethane (anhydrous), toluene (anhydrous), potassium carbonate (K₂CO₃), calcium chloride (CaCl₂) and magnesium sulfate (MgSO₄) purchased from Sigma-Aldrich (MO, USA), ethyl ether (anhydrous) purchased from Thermo Fisher Scientific (MA, USA), micro-filtered and lyophilized sodium alginate (MVG GRGDSP-coupled; high G ratio; high M_w) purchased from NovaMatrix (Dupont, DE, USA), high purity chitosan (99% degree of deacetylation; M_w: 100 kDa), as primers for tissues, purchased from Sigma-Aldrich (MO, USA), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and sulfated N-hydroxysuccinimide (Sulfo-NHS), as coupling reagents for tissue and device primers, purchased from Thermo Fisher Scientific (MA, USA). The solvents included ultrapure distilled water, phosphate-buffered saline (PBS; pH 7.4), and 2-(N-morpholino)ethanesulfonic acid (MES; pH 5.0) purchased from Thermo Fisher Scientific (MA, USA). All chemicals were used as received. Tissues and organs for bench studies were purchased from Animal Biotech Industries (ABI; PA, USA) and were cleaned with soap and water before adhesion measurements.

Yang Q., Wei T., Yin R.T., Wu M., Xu Y., Koo J., Choi Y.S., Xie Z., Chen S.W., Kandela I., Yao S., Deng Y., Avila R., Liu T.L., Bai W., Yang Y., Han M., Zhang Q., Haney C.R., Lee K.B., Aras K., Wang T., Seo M.H., Luan H., Lee S.M., Brikha A., Ghoreishi-Haack N., Tran L., Stepien I., Aird F., Waters E.A., Yu X., Banks A., Trachiotis G.D., Torkelson J.M., Huang Y., Kozorovitskiy Y., Efimov I.R, & Rogers J.A. (2021). Photocurable bioresorbable adhesives as functional interfaces between flexible bioelectronic devices and soft biological tissues. Nature materials, 20(11), 1559-1570.

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Quantification of 5-MeO-MiPT in Biological Samples

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The reference standard of 5-MeO-MiPT was bought from Lipomed Inc (Cambridge, MA). The internal standard, diazepam-d5 10 μg/mL in methanol (MeOH), was purchased from Restek Corporation (Bellefonte, PA). High-performance liquid chromatography (HPLC)-grade MeOH and water, sodium chloride (NaCl), acetone, dimethyl sulfoxide (DMSO), ammonium hydroxide, ethyl acetate, and ultrapure MeOH were bought from Merck KGaA (Darmstadt, Germany). Ammonium formate and phosphate buffer (0.1 M, pH 4.4) was purchased from EMD Chemicals, Inc (Gibbstown, NJ). Formaldehyde, ethanol, and hydrogen peroxide (H₂O₂) were bought from Merck KGaA (Darmstadt, Germany). Phosphate-buffered saline (PBS; pH 7.4) was purchased from Thermo Fisher Scientific (Waltham, MA). Hematoxylin and eosin (H&E) were purchased from Sigma-Aldrich (Darmstadt, Germany). Trypsinized (2% trypsin in 50 mM Tris buffer) and 3,3′-diaminobenzidine (DAB; DeadEnd Colorimetric TUNEL System) were purchased from Abcam (Cambridge, UK). Caspase-3 (1:100, sc-7272) and caspase-8 (1:100, sc-56070) were purchased from Santa Cruz Biotechnology Inc (Santa Cruz, CA). Biotinylated secondary antibody and streptavidin-bound horseradish peroxidase were purchased from Zymed Laboratories Inc (South San Francisco, CA) (Histostain Plus Kit; 85–9043). Mayer’s hematoxylin was purchased from Zymed Laboratories Inc (South San Francisco, CA).

Altuncı Y.A., Aydoğdu M., Açıkgöz E., Güven Ü., Düzağaç F., Atasoy A., Dağlıoğlu N, & Akgür S.A. (2021). New Psychoactive Substance 5-MeO-MiPT In vivo Acute Toxicity and Hystotoxicological Study. Balkan Medical Journal, 38(1), 34-42.

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SARS-CoV-2 Spike RBD Protein Binding Assay

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Phosphate-buffered saline (PBS; pH 7.4) was purchased from Thermo Fisher Scientific, Inc. (USA). Albumin from bovine serum (BSA) was purchased from FUJIFILM Wako Pure Chemical Corporation (Japan). Recombinant SARS-CoV-2 spike RBD protein (39 kDa) was purchased from SignalChem Biotech, Inc. (Canada). Anti SARS-CoV-2 Spike RBD polyclonal rabbit IgG antibody was purchased from Sino Biological, Inc. (China). Anti-avian influenza A hemagglutinin rabbit IgG antibody was purchased from Abcam, Inc. (USA). Healthy human serum was purchased from Cosmo Bio Co., Ltd., (Japan). COVID-19 positive and negative human serum samples were purchased from Raybiotech, Inc. (USA). Positive serum samples were collected from patients who were positive for COVID-19 by PCR or antigen testing, and have been inactivated. Polydimethylsiloxane (PDMS; SILPOT 184 W/C) was purchased from Dow Corning Toray Co., Ltd. (Japan). A negative photoresist (SU-8 3050) was purchased from Nippon Kayaku Co., Ltd. (Japan). HiLyte Fluor™ 647 Labeling Kit-NH₂ were purchased from Dojindo Molecular Technologies, Inc. (Japan). Two COVID-19 IgM/IgG rapid test LFIA kits, kit A and kit B, were purchased from GenBody Inc. (Korea) and Epigentek (USA), respectively.

Nishiyama K., Takahashi K., Fukuyama M., Kasuya M., Imai A., Usukura T., Maishi N., Maeki M., Ishida A., Tani H., Hida K., Shigemura K., Hibara A, & Tokeshi M. (2021). Facile and rapid detection of SARS-CoV-2 antibody based on a noncompetitive fluorescence polarization immunoassay in human serum samples. Biosensors & Bioelectronics, 190, 113414.

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PEG-Collagen Hydrogel Characterization

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Unless otherwise noted, all chemicals and solvents were of analytical grade and used as advised by the manufacturer. 4 and 8-arm PEG succinimidyl glutarate (PEG) was purchased from JenKem Technology (Texas, USA). Collagenase, dimethyl sulfoxide (DMSO), fluorescamine, sodium hydroxide solution (1.0 N), bovine serum albumin (BSA), fibronectin, cholera subunit A, insulin, Triton-X, and Cell Counting kit-8 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Phosphate-buffered saline (PBS) pH 7.4, 10X PBS, Slide-A-Lyzer dialysis kit (3.5 kDa MWCO), collagen I bovine protein solution (5 mg/mL), Collagenase, Dulbecco’s modified eagle medium nutrient mixture F-12 (DMEM/F-12) with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), insulin-transferrinselenium (ITS), Dulbecco’s phosphate-buffered saline (DPBS), antibiotic-antimycotic, Live/Dead viability/cytotoxicity staining kit, and paraformaldehyde (PFA) were purchased from Thermo Fisher Scientific (Massachusetts, USA).

Na K.S., Fernandes-Cunha G.M., Varela I.B., Lee H.J., Seo Y.A, & Myung D. (2021). Effect of mesenchymal stem cells encapsulated within in situ forming PEG-collagen hydrogels on alkali-burned corneas in an ex vivo organ culture model. Cytotherapy, 23(6), 500-509.

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