Fatty-acid-free human serum albumin (HSA) was purchased from Sigma-Aldrich and used without purification. Stock solutions of HSA were prepared in tris-HCl buffer at pH 7.4 with 0.10 M NaCl. Apixaban (AP), rivaroxaban (RV), and betrixaban (BE) were purchased from Ambeed Inc. Edoxaban (ED). Edoxaban tosylate hydrate (EDT) and betrixaban maleate (BEM) were purchased from AKScientific. Stock solutions of AP, RV, ED, BE, EDT, and BEM were prepared in DMSO. For the complexation studies using HSA and the anticoagulants, the added DMSO was less than 2% [28 (link)].
Fatty acid free human serum albumin
Manufactured by Merck Group
Sourced in United States
Fatty acid-free human serum albumin is a laboratory product that serves as a highly purified form of the blood protein albumin, with the fatty acids removed. It is commonly used as a component in cell culture media and other in vitro applications that require a serum-derived protein supplement.
Automatically generated - may contain errors
Lab products found in correlation
Sodium phosphate monobasic, by Thermo Fisher Scientific (2 mentions)
Sodium dodecyl sulfate (sds), by Thermo Fisher Scientific (2 mentions)
Sodium phosphate dibasic anhydrous, by Thermo Fisher Scientific (2 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (2 mentions)
Acetone, by Thermo Fisher Scientific (2 mentions)
3 4 morpholino propane sulfonic acid mops, by Thermo Fisher Scientific (2 mentions)
4 2 hydroxyethyl 1 piperazineethanesulfonic acid hepes, by Thermo Fisher Scientific (2 mentions)
Fatty acid free bovine serum albumin, by Merck Group (2 mentions)
1 2 distearoyl sn glycero 3 phosphoethanolamine n methoxy polyethylene glycol 2000 dspe peg, by Avanti Polar Lipids (2 mentions)
Midazolam, by Fujifilm (1 mentions)
Phenylephrine, by Fujifilm (1 mentions)
Eletriptan, by Merck Group (1 mentions)
Sumatriptan, by Tokyo Chemical Industry (1 mentions)
Sertraline, by Fujifilm (1 mentions)
Mao a, by Corning (1 mentions)
Sample buffer, by Bio-Rad (1 mentions)
Mouse albumin, by Merck Group (1 mentions)
Human alpha1 acid glycoprotein, by Merck Group (1 mentions)
Seeblue plus2, by Thermo Fisher Scientific (1 mentions)
Tris glycine gel, by Bio-Rad (1 mentions)
11 protocols using fatty acid free human serum albumin
1
Anticoagulant-albumin Complexation Studies
2
Liver Microsome Preparation and Characterization
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Midazolam, rizatriptan, phenylephrine and sertraline were purchased from Wako Pure Chemical Industries (Osaka, Japan). Sumatriptan and S-citalopram were purchased from Tokyo Chemical Industry (Tokyo, Japan). Imipramine was purchased from Nakalai Tesque (Kyoto, Japan). Eletriptan and fatty acid-free human serum albumin (HSA) were obtained from Sigma-Aldrich (St. Louis, MO). All other chemicals and reagents were analytical-grade products from commercial sources. Pooled HLMs (50 donors, #PLo50BA), CD-1 mouse liver microsomes, SD rat liver microsomes and beagle dog liver microsomes were purchased from Thermo Fisher Scientific (Waltham, MA). Pooled HLMt (5 donors, #1110152) was purchased from Sekisui XenoTech.
(Kansas City, KS). An MAO expression system (Supersomes: MAO-A, MAO-B and control) was purchased from Corning (Corning, NY). Individual HLMs and HLMt were prepared according to the previously reported method (13) . Human liver blocks were obtained from Human and Animal Bridging Research Organization (Chiba, Japan). The study protocol was approved by the ethical committees of the School of Medicine in Kanazawa University. Individual HLMs and HLMt were independently prepared to prevent loss derived from sequential purification.
(Kansas City, KS). An MAO expression system (Supersomes: MAO-A, MAO-B and control) was purchased from Corning (Corning, NY). Individual HLMs and HLMt were prepared according to the previously reported method (13) . Human liver blocks were obtained from Human and Animal Bridging Research Organization (Chiba, Japan). The study protocol was approved by the ethical committees of the School of Medicine in Kanazawa University. Individual HLMs and HLMt were independently prepared to prevent loss derived from sequential purification.
3
APC Compound Binding Assay in Plasma
4
Preparation of Prodan-Conjugated Albumin
5
PEG-Polymer Characterization Protocols
6
Rapid Equilibrium Dialysis Protocol
7
Deuterium-Labeled Fatty Acid Incorporation in Erythrocytes
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Human peripheral blood erythrocytes from healthy human volunteers were isolated by centrifugation and aspiration of the platelet-rich plasma and buffy coat layer. Erythrocytes were purified by resuspension in RPMI 1640 (Lonza) [10% hematocrit (ht)] and centrifugation followed by aspiration of the top 10% of the erythrocyte layer (this purification procedure was carried out with five repetitions). Purified erythrocytes were then resuspended (20% ht) in RPMI 1640 containing deuterium-labeled EPA and DHA (d5-EPA, 10 μM and d5-DHA, 10 μM) with fatty acid–free human serum albumin (Sigma-Aldrich) and were incubated for 3 hours at 37°C to allow maximal incorporation of fatty acids into phospholipid membranes [see (64 (link)) for further details]. Cells were washed five times with 10 volumes of RPMI 1640 to remove extracellular free fatty acids.
8
Platelet Function Assay with Collagen
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Fatty acid–free human serum albumin and tetramethylrhodamine (TRITC) phalloidin were provided by Sigma‐Aldrich (St. Louis, MO, USA). Fibrinogen was from Fresenius Kabi (Lake Zurich, IL, USA). Horm fibrillar type‐I collagen from equine Achilles tendon, used to coat flow chambers (200 μg/ml) was purchased from Takeda (Tokyo, Japan). ReoPro was from Eli Lilly (Indianapolis, IN, USA). The anti‐GPVI blocking antibody, 1G5, was developed by Elizabeth Gardiner (ANU, Australia). IV.3 was from Stemcell Technologies (Vancouver, BC, Canada). The mouse IgG2b use as a negative control (clone MPC‐11) was from Merck‐Millipore (Molsheim, France). RAM‐1 is an anti‐GPIbβ antibody developed at U1255 (François Lanza). Recombinant hirudin, used as an anticoagulant to directly block thrombin (100 U/mL), was from Transgene (Illkirch‐Graffenstaden, France). 3,3’‐dihexyloxacarbocyanine iodide (DIOC6) was from Molecular Probes (Paisley, UK). Paraformaldehyde was from VWR (Strasbourg, France).
9
Protein-Polymer Interaction Protocols
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Poly(ethylene glycol) with a molecular weight of 2000 g/mol (PEG2000) was obtained from Fluka. Poly(ethylene glycol-block-caprolactone) (PEG-CL; AK073; 70411STR-B) and poly(ethylene glycol-blockpolylactide) (PEG-LA; AK009; 190813RAI-B) were obtained from Polyscitech and used for BSA ITC and CD experiments. Poly(ethylene glycol-block-caprolactone) (PEG-CL; AK073; 40319SMS-A) and poly(ethylene glycol-block-polylactide) (PEG-LA; AK009; 180329YSK-A) were obtained from Polyscitech and used for protein fluorescence and HSA ITC and CD experiments. 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (PEG-DSPE) was obtained from Avanti Polar Lipids. Fatty acid-free bovine serum albumin (BSA) was obtained from Millipore Sigma. Fatty acid-free human serum albumin and pyrene were obtained from Sigma-Aldrich. Acetone, sodium chloride, sodium phosphate dibasic anhydrous, sodium phosphate monobasic, 3-(4-morpholino)propane sulfonic acid (MOPS), 4-(2-hydroxyethyl)-1piperazineethanesulfonic acid (HEPES), and sodium dodecyl sulfate (SDS) were obtained from Fisher Scientific. Regenerated cellulose dialysis membranes with a molecular weight cutoff of 3500 g/mol were obtained from Spectrum Laboratories.
10
Peptide Synthesis with Diverse Reagents
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NovaSyn TGA resin (amine density of 0.2 mmol/g), Fmoc-Glu(Odmab)-OH, Fmoc-Asp(OtBu)-OH and Fmoc-Gly-OH were obtained from Novabiochem, Merck (Tokyo, Japan). N-[1-(Cyano-2-ethoxy-2-oxoethylideneaminooxy) dimethylamino (morpholino)] uronium hexafluorophosphate (COMU), Fmocβ-Ala-OH, Fmoc-Glu-OtBu, diisopropylethylamine (DIPEA), dichloromethane (DCM), 1-methyl-2-pyrrolidone (NMP), and piperidine were purchased from Watanabe Chemical (Tokyo, Japan).
Pyridine, N,N-dimethyl-4-aminoPyridine benzoic anhydride (DMAP) and N,N-diisopropylcarbodiimide (DIC) were obtained from Wako Pure Chemicals (Osaka, Japan). Boc-11-amino-3,6,9-trioxaundecanoic acid•DCHA (Boc-mini-PEG3) and Fmoc-11-3,6,9-trioxaundecanoic acid (Fmoc-mini-PEG3) were purchased from Peptides International (Kentucky, USA). N,N-Dimethylformamide (DMF) was purchased from Kanto Chemical (Tokyo, Japan). All reagents were used without further purification. Fatty acid free human serum albumin and myristic acid were purchased from Sigma-Aldrich. Tetradecylamine, hexadecylamine and sodium salt of 8-anilino-1-naphthalenesulfonate (ANS•Na) were purchased from Tokyo Chemical Industry (Tokyo, Japan).
Pyridine, N,N-dimethyl-4-aminoPyridine benzoic anhydride (DMAP) and N,N-diisopropylcarbodiimide (DIC) were obtained from Wako Pure Chemicals (Osaka, Japan). Boc-11-amino-3,6,9-trioxaundecanoic acid•DCHA (Boc-mini-PEG3) and Fmoc-11-3,6,9-trioxaundecanoic acid (Fmoc-mini-PEG3) were purchased from Peptides International (Kentucky, USA). N,N-Dimethylformamide (DMF) was purchased from Kanto Chemical (Tokyo, Japan). All reagents were used without further purification. Fatty acid free human serum albumin and myristic acid were purchased from Sigma-Aldrich. Tetradecylamine, hexadecylamine and sodium salt of 8-anilino-1-naphthalenesulfonate (ANS•Na) were purchased from Tokyo Chemical Industry (Tokyo, Japan).
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