SP2/0 mouse myeloma cell line (kindly provided by Dr. L. Wysocki, University of Colorado at Denver, Denver, CO, USA) and previously generated B-cell hybridomas (4H4, 4G10, 2A10 and 1F8) were cultured as already described [20 (link)]. In detail, mouse cell lines were maintained in B cell medium (BCM) consisting of RPMI 1640 medium (Sigma-Aldrich, Saint Louis, MO, USA) containing 10% FBS (Atlanta Biologicals), 23.8 mM sodium bicarbonate (Thermo Fisher Scientific, Waltham, MA, USA), 7.5 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES; Amresco, Solon, OH, USA), 170 mM penicillin G (Tokyo Chemical Industry, Tokyo, Japan), 137 mM streptomycin (Sigma-Aldrich), 50 mM 2-mercaptoethanol (Sigma-Aldrich), nonessential amino acids (Thermo Fisher Scientific) and 1 mM sodium pyruvate (Thermo Fisher Scientific). For monoclonal antibody (mAb) production, hybridoma cell lines were grown in BCM containing 2% Super Low IgG FBS (HyClone, Logan, UT, USA).
4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes
Manufactured by Avantor
Sourced in United States
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) is a zwitterionic organic chemical compound used as a buffering agent in biological research and cell culture applications. It is a common component in various cell culture media and is used to maintain a stable pH environment for cells.
Automatically generated - may contain errors
Lab products found in correlation
Sodium bicarbonate, by Thermo Fisher Scientific (3 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Penicillin g, by Tokyo Chemical Industry (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
2 mercaptoethanol, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (1 mentions)
Chrysin, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Avantor (1 mentions)
Dibenzylfluorescein, by Merck Group (1 mentions)
Thiazolyl blue tetrazolium blue mtt, by Merck Group (1 mentions)
Ultra clear centrifuge tube, by Beckman Coulter (1 mentions)
Quercitrin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Quercetin, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetic acid, by RCI Labscan (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Potassium persulfate, by Merck Group (1 mentions)
7 protocols using 4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes
1
Culturing Myeloma and Hybridoma Cell Lines
2
Biochemical Assay Reagents Procurement
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Chrysin, quercetin, quercitrin, dibenzylfluorescein, and thiazolyl blue tetrazolium blue (MTT) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Potassium chloride (KCl) and methanol were obtained from Merck (Darmstadt, Germany). Acetic acid was obtained from RCI Labscan (Bangkok, Thailand). 4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was purchased from AMRESCO (Solon, OH, USA). Nicotinamide adenine dinucleotide phosphate (NADPH) was obtained from Tocris (Ellisville, MO, USA). Antibiotic-antimycotic Dulbecco's modified Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco BRL Life Technologies (Grand Island, NY, USA). Dimethyl sulfoxide (DMSO) was obtained from Amresco (Salon, OH, USA). All solvents were of analytical grade. Ninety-six-well plates were obtained from Costar (Cambridge, MA, USA). Ultra-clear centrifuge tubes were purchased from Beckman Coulter Inc. (Brea, CA, USA).
3
Synthesis of Stimuli-Responsive Polymeric Nanoparticles
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All chemicals, if not stated otherwise, were used as provided by the supplier. Triton X-45, acrylic acid (AA), methacrylic acid (MAA), N, N’ methylenebis(acrylamide) (BIS), 2,2,2-trifluoroethyl methacrylate (TFMA), sodium dodecyl sulfate (SDS), potassium persulfate (KPS), BODIPY, and Nile red were purchased from Sigma-Aldrich. N-Isopropylacrylamide (NIPAM) was purchased from Sigma-Aldrich and purified by recrystallization in Toluene/Hexane 50:50. 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was purchased from VWR.
4
Glycosyltransferase Activity Assay
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Reagent grade chloroform, methanol, and silica
gel 60 thin-layer chromatography (TLC) plates (layer thickness of
150–200
μm) were obtained from EMD Chemicals Inc. (Gibbstown,
NJ). [32P]Pi, UDP-[U-14C]glucose,
UDP-[U-14C]galactose, and GDP-[U-14C]mannose
were from
PerkinElmer Life and Analytical
Sciences, Inc. (Waltham, MA). Yeast extract, agar, and tryptone were
from BD Biosciences. Sodium chloride and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES) were from VWR International (West Chester, PA). Triton
X-100 was from Thermo Fisher Scientific (Waltham, MA). Isopropyl 1-thio-β-d -galactopyranoside (IPTG) was from Invitrogen Corp. (Carlsbad,
CA). UDP-glucose, UDP-galactose, UDP-glucuronic acid, UDP-galacuronic
acid, ADP-glucose, GDP-glucose, UDP-N-acetylglucosamine,
and ADP-mannose were purchased from Sigma-Aldrich.
The P-10 desalting column was from GE. All other chemicals were reagent
grade and were purchased from either Sigma-Aldrich (St. Louis, MO)
or Mallinckrodt Baker, Inc. (Phillipsburg, NJ).
gel 60 thin-layer chromatography (TLC) plates (layer thickness of
150–200
μm) were obtained from EMD Chemicals Inc. (Gibbstown,
NJ). [32P]Pi, UDP-[U-14C]glucose,
UDP-[U-14C]galactose, and GDP-[U-14C]mannose
were from
PerkinElmer Life and Analytical
Sciences, Inc. (Waltham, MA). Yeast extract, agar, and tryptone were
from BD Biosciences. Sodium chloride and 4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid (HEPES) were from VWR International (West Chester, PA). Triton
X-100 was from Thermo Fisher Scientific (Waltham, MA). Isopropyl 1-thio-β-
CA). UDP-glucose, UDP-galactose, UDP-glucuronic acid, UDP-galacuronic
acid, ADP-glucose, GDP-glucose, UDP-N-acetylglucosamine,
and ADP-mannose were purchased from Sigma-Aldrich.
The P-10 desalting column was from GE. All other chemicals were reagent
grade and were purchased from either Sigma-Aldrich (St. Louis, MO)
or Mallinckrodt Baker, Inc. (Phillipsburg, NJ).
5
Platelet Activation and Fibrinogen Binding
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Adenosine diphosphate (ADP) was from Sigma-Aldrich (Zwijndrecht, the Netherlands). Allophycocyanin (APC)-conjugated monoclonal Mouse Anti-Human P-selectin (CD62P) antibody clone AK4, Phycoerythrin (PE) conjugated monoclonal Mouse Anti-Human P-selectin antibody clone AK4, BD FACSCanto II, and FACSCanto II Diva software version 8.0.1 were from BD Biosciences (Franklin Lakes, New Jersey, USA). Fluorescein isothiocyanate (FITC) conjugated polyclonal Rabbit Anti-Human fibrinogen antibody (F011102-2) was from Dako (now Agilent, Santa Clara, CA, USA).
Formaldehyde (37%) was from Calbiochem (San Diego, California, USA). MgSO4 was from Merck (Darmstadt, Germany). NaCl, KCl were from Sigma–Aldrich (St. Louis, MO, USA). PAR (protease-activated receptor)-1 agonist SFLLRN (TRAP-6) was from Bachem (Bubenhof, Zwitserland). 1.2-mL polypropylene tubes were obtained from BRAND GmbH & Co. KG (Wertheim, Germany). 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was from VWR International (Amsterdam, The Netherlands). Ninety-six-well PS flat-bottom plates were from Greiner Bio-one (Alphen aan den Rijn, The Netherlands).
Formaldehyde (37%) was from Calbiochem (San Diego, California, USA). MgSO4 was from Merck (Darmstadt, Germany). NaCl, KCl were from Sigma–Aldrich (St. Louis, MO, USA). PAR (protease-activated receptor)-1 agonist SFLLRN (TRAP-6) was from Bachem (Bubenhof, Zwitserland). 1.2-mL polypropylene tubes were obtained from BRAND GmbH & Co. KG (Wertheim, Germany). 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) was from VWR International (Amsterdam, The Netherlands). Ninety-six-well PS flat-bottom plates were from Greiner Bio-one (Alphen aan den Rijn, The Netherlands).
6
Hydrophobic Coating Nanoparticle Synthesis
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Glaco Mirror Coat Zero™ was purchased from Soft99 (Osaka, Japan). It is composed of silica nanoparticles suspension (hydrophobically modified with alkyl groups) dispersed in isopropanol. The nanoparticle concentration in the Glaco™ solution is about 4 g.L -1 , as quantified by weighing the dry residue after evaporation of the solvent. Ultrapure grade water (18 MΩ.cm -1 ) was produced on a Milli-Q equipment (Merck Millipore, USA). Sodium hydroxide (NaOH) and N,Ndimethylformamide (DMF, 99%) were purchased from Sigma-Aldrich (St Quentin Fallavier, France). 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (HEPES) was purchased from VWR (Fontenay-sous-bois, France). Brilliant Blue FCF (food coloring, E133) from Vahiné (Avignon, France) was used as an aqueous colorant.
Mesityl Oxide (4-metyl-4-penten-2-one) was purchased from Alfa Aesar (Thermo Fischer Scientific, Karlsruhe, Germany).
Mesityl Oxide (4-metyl-4-penten-2-one) was purchased from Alfa Aesar (Thermo Fischer Scientific, Karlsruhe, Germany).
7
Protein and Cellulose Nanocrystal Preparation
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HEWL (Mw = 14.3 kDa, purity > 90%) was purchased from Sigma-Aldrich. Purified BLG was (Mw = 18.4 kDa, purity > 97%) provided by the Food and Bioprocess Engineering group at the Technical University of Munich [46] (link). CNCs were purchased from CelluForce. TO-CNFs were prepared in-house according to a previously published protocol [41] (link).
The 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, purity > 99.5%), sodium hydroxide (NaOH, purity > 99%) and hydrochloric acid (HCl, 37%) were obtained from VWR. The (3-aminopropyl)triethoxysilane (APTES, purity > 98%) was purchased from Sigma-Aldrich. All chemicals were used as received without further purification.
The 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES, purity > 99.5%), sodium hydroxide (NaOH, purity > 99%) and hydrochloric acid (HCl, 37%) were obtained from VWR. The (3-aminopropyl)triethoxysilane (APTES, purity > 98%) was purchased from Sigma-Aldrich. All chemicals were used as received without further purification.
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