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Avantor hepes
Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
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Images

1) Product Images from "Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug"

Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

Journal: Pharmaceutical Research

doi: 10.1007/s11095-016-2002-5

Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
Figure Legend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.

Techniques Used: Incubation

Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.
Figure Legend Snippet: Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.

Techniques Used: Incubation, Generated

Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.
Figure Legend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.

Techniques Used: Incubation

2) Product Images from "Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris"

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris

Journal: Protein expression and purification

doi: 10.1016/j.pep.2009.10.011

D-glucose transport activity of purified GLUT1 reconstituted into proteoliposomes Proteoliposomes were prepared by gel filtration through Sephadex G-50. 12.5 µg of soybean phospholipids in 20 mM HEPES, 0.30 M NaCl were sonicated, adjusted to 37 mM CHAPS, incubated for 2 hours at 25 °C, and then 12.5 µg/ml of purified aglyco-GLUT1 was added. Two hundred microliters were filtered twice through a 2 ml bed of pre-spun Sephadex G-50 equilibrated with 20 mM HEPES, 0.15 M NaCl. Proteoliposomes (27 ul) were added to 3 µl of uptake solution (0.2 mM L- or D- [ 3 H] glucose, final concentration) and the uptake was quenched by the addition of 1 µl of 60 mM HgCl 2 . This mix was filtered through spin columns of 0.6 ml Sephadex G-50 pre-equilibrated with 20 mM HEPES, 0.15 M NaCl, 2 mM HgCl 2 . Each point is the average of 3 independent measurements.
Figure Legend Snippet: D-glucose transport activity of purified GLUT1 reconstituted into proteoliposomes Proteoliposomes were prepared by gel filtration through Sephadex G-50. 12.5 µg of soybean phospholipids in 20 mM HEPES, 0.30 M NaCl were sonicated, adjusted to 37 mM CHAPS, incubated for 2 hours at 25 °C, and then 12.5 µg/ml of purified aglyco-GLUT1 was added. Two hundred microliters were filtered twice through a 2 ml bed of pre-spun Sephadex G-50 equilibrated with 20 mM HEPES, 0.15 M NaCl. Proteoliposomes (27 ul) were added to 3 µl of uptake solution (0.2 mM L- or D- [ 3 H] glucose, final concentration) and the uptake was quenched by the addition of 1 µl of 60 mM HgCl 2 . This mix was filtered through spin columns of 0.6 ml Sephadex G-50 pre-equilibrated with 20 mM HEPES, 0.15 M NaCl, 2 mM HgCl 2 . Each point is the average of 3 independent measurements.

Techniques Used: Activity Assay, Purification, Filtration, Sonication, Incubation, Concentration Assay

Related Articles

Centrifugation:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C. .. Two hundred microliters of the proteoliposome mixture were loaded onto a 2 ml column of pre-spun Sephadex G-50 equilibrated with 20 mM HEPES, 0.15 M NaCl and the centrifugation-filtration was conducted twice in succession.

Article Title: A Maltose-Binding Protein Fusion Construct Yields a Robust Crystallography Platform for MCL1
Article Snippet: .. Protein Purification of MCL1, MBP-MCL1 Fusion, TRX-MCL1 Fusion and Mutants Cells were resuspended in 50mM HEPES pH 7.5, 500mM NaCl, 5% glycerol, 0.001% 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) (JT Baker), 0.5mM Tris-(2-carboxyethyl)phosphine (TCEP) (VWR), 250 U Benzonase (Novagen), 100 mg lysozyme (Sigma) and one complete EDTA free protease inhibitor tablet (Roche), lysed via sonication and clarified via centrifugation. ..

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. ProteinG beads were collected by brief centrifugation and washed 3 times in NP40 lysis buffer.

Luciferase:

Article Title: Engineering triiodothyronine (T3) nanoparticle for use in ischemic brain stroke
Article Snippet: HEPES, NaCl, and EDTA were received from VWR International (West Chester, PA, USA). .. Dual luciferase kit was from Promega (Madison, WI, USA).

Filtration:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: Proteoliposomes were prepared by gel filtration [ ] using Sephadex G-50. .. 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C.

Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug
Article Snippet: .. The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)). ..

Article Title: Phosphatidic acid mediates the targeting of tBid to induce lysosomal membrane permeabilization and apoptosis [S]
Article Snippet: Experiments were performed in a Superdex-200 (1.5 × 45 cm) column equilibrated with 100 mM KCl, 10 mM HEPES, and 0.2 mM EDTA (pH 7.0) with or without 2% (w/v) CHAPS (J. T. Baker Inc.) at a 1 ml/min flow rate. .. The column was calibrated using protein gel filtration standards (Bio-Rad).

Construct:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Recombinant gH-V5/gL-Flag complexes were precipitated from the lysates of 293T cells transfected with the respective expression constructs. .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

Enzyme-linked Immunosorbent Assay:

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: .. Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

Incubation:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: .. 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C. .. 12.5 µg/ml of purified aglyco- GLUT1 was then added to the lipids.

Article Title: Potential adverse effects of botanical supplementation in high-fat-fed female mice
Article Snippet: Analysis of protein expression Skeletal muscle and liver lysates were prepared from powdered tissue by homogenizing in 25 mM HEPES, pH 7.4, 1% Igepal CA630, 137 mM NaCl, 1 mM PMSF, 10 μg/ml aprotinin, 1 μg/ml pepstatin, 5 μg/ml leupeptin, 10 mM Na4 P2 O7 , 100 mM NaF, and 2 mM NaVO4 using a Sonifier 450 homogenizer (VWR, Radnor, PA). .. Nitrocellulose membranes were incubated with antibodies for 1–2 h at room temperature or overnight at 4 °C as indicated.

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. Amount of input gH/gL between mutants was normalized by diluting lysates with cell lysate from non-transfected 293T cells prior to immunoprecipitation according to Western blot analysis of lysates and evaluation of the gH/gL content for each construct.

Expressing:

Article Title: Potential adverse effects of botanical supplementation in high-fat-fed female mice
Article Snippet: .. Analysis of protein expression Skeletal muscle and liver lysates were prepared from powdered tissue by homogenizing in 25 mM HEPES, pH 7.4, 1% Igepal CA630, 137 mM NaCl, 1 mM PMSF, 10 μg/ml aprotinin, 1 μg/ml pepstatin, 5 μg/ml leupeptin, 10 mM Na4 P2 O7 , 100 mM NaF, and 2 mM NaVO4 using a Sonifier 450 homogenizer (VWR, Radnor, PA). .. Protein concentrations were determined using a BCA assay (Thermo Fisher Scientific, Rockford, IL) according to the manufacturer’s instructions.

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Recombinant gH-V5/gL-Flag complexes were precipitated from the lysates of 293T cells transfected with the respective expression constructs. .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

BIA-KA:

Article Title: Potential adverse effects of botanical supplementation in high-fat-fed female mice
Article Snippet: Analysis of protein expression Skeletal muscle and liver lysates were prepared from powdered tissue by homogenizing in 25 mM HEPES, pH 7.4, 1% Igepal CA630, 137 mM NaCl, 1 mM PMSF, 10 μg/ml aprotinin, 1 μg/ml pepstatin, 5 μg/ml leupeptin, 10 mM Na4 P2 O7 , 100 mM NaF, and 2 mM NaVO4 using a Sonifier 450 homogenizer (VWR, Radnor, PA). .. Analysis of protein expression Skeletal muscle and liver lysates were prepared from powdered tissue by homogenizing in 25 mM HEPES, pH 7.4, 1% Igepal CA630, 137 mM NaCl, 1 mM PMSF, 10 μg/ml aprotinin, 1 μg/ml pepstatin, 5 μg/ml leupeptin, 10 mM Na4 P2 O7 , 100 mM NaF, and 2 mM NaVO4 using a Sonifier 450 homogenizer (VWR, Radnor, PA).

Modification:

Article Title: Engineering triiodothyronine (T3) nanoparticle for use in ischemic brain stroke
Article Snippet: DMEM (Dulbecco’s modified eagle medium), fetal calf serum, and penicillin–streptomycin were purchased from Hyclone (Logan, UT, USA). .. HEPES, NaCl, and EDTA were received from VWR International (West Chester, PA, USA).

Western Blot:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Paragraph title: Immunoprecipitation and Western blot analysis ... Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

High Performance Liquid Chromatography:

Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug
Article Snippet: .. The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)). ..

Flow Cytometry:

Article Title: Probing Prokaryotic Social Behaviors with Bacterial "Lobster Traps"
Article Snippet: .. Structures were fabricated using 400 mg ml−1 BSA (Equitech-Bio) and 8.5 mM rose bengal (Sigma) as a photosensitizer in 20 mM HEPES–0.1 M NaCl buffer (pH 7.4) on no. 1 coverglass (VWR) within a single-pass flow cell system ( ). ..

Article Title: Phosphatidic acid mediates the targeting of tBid to induce lysosomal membrane permeabilization and apoptosis [S]
Article Snippet: .. Experiments were performed in a Superdex-200 (1.5 × 45 cm) column equilibrated with 100 mM KCl, 10 mM HEPES, and 0.2 mM EDTA (pH 7.0) with or without 2% (w/v) CHAPS (J. T. Baker Inc.) at a 1 ml/min flow rate. .. The column was calibrated using protein gel filtration standards (Bio-Rad).

Concentration Assay:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C. .. Proteoliposomes (27 µl) were added to 3 µl of uptake solution (0.2 mM L- or D- [3 H] glucose, final concentration), incubated for various time periods, and the uptake was then stopped by the addition of 1 µl of 60 mM HgCl2 .

Protease Inhibitor:

Article Title: A Maltose-Binding Protein Fusion Construct Yields a Robust Crystallography Platform for MCL1
Article Snippet: .. Protein Purification of MCL1, MBP-MCL1 Fusion, TRX-MCL1 Fusion and Mutants Cells were resuspended in 50mM HEPES pH 7.5, 500mM NaCl, 5% glycerol, 0.001% 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) (JT Baker), 0.5mM Tris-(2-carboxyethyl)phosphine (TCEP) (VWR), 250 U Benzonase (Novagen), 100 mg lysozyme (Sigma) and one complete EDTA free protease inhibitor tablet (Roche), lysed via sonication and clarified via centrifugation. ..

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. Amount of input gH/gL between mutants was normalized by diluting lysates with cell lysate from non-transfected 293T cells prior to immunoprecipitation according to Western blot analysis of lysates and evaluation of the gH/gL content for each construct.

Northern Blot:

Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug
Article Snippet: Materials Liposomes containing 50 mg/ml ciprofloxacin (expressed in terms of ciprofloxacin hydrochloride) in a pH 6.0 histidine buffer were manufactured by Northern Lipids Inc. (Burnaby, BC, Canada) and Sigma-Tau (Indianapolis, IN). .. The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

Cell Culture:

Article Title: The extended recovery ring-stage survival assay provides a superior association with patient clearance half-life and increases throughput
Article Snippet: .. Parasite culture Plasmodium falciparum isolates were cultured using standard methods in human red blood cells (RBC) (Biochemed Services, Winchester, VA and Interstate Blood Bank, Memphis, TN) suspended in complete medium (CM) containing RPMI 1640 with l -glutamine (Gibco, Life Technologies.), 50 mg/L hypoxanthine (Calbiochem, Sigma-Aldrich), 25 mM HEPES (Corning, VWR, 0.5% Albumax II (Gibco, Life Technologies.), 10 mg/L gentamicin (Gibco, Life Technologies) and 0.225% NaHCO3 (Corning, VWR) at 5% haematocrit. ..

other:

Article Title: Augmenting Sulfur Metabolism and Herbivore Defense in Arabidopsis by Bacterial Volatile Signaling
Article Snippet: Supernatant aliquots were neutralized with 200 mM HEPES (pH 12.4), reduced with dithiothreitol and sulfhydryl groups derivatized with monobromobimane (VWR).

Article Title:
Article Snippet: HEPES was obtained from J. T. Baker (Mallinckrodt Baker, Inc., Phillipsburg, NJ).

Article Title: Selectivity of ABT-089 for ?4?2* and ?6?2* nicotinic acetylcholine receptors in brain
Article Snippet: Sucrose was obtained from Fisher Chemical Co., Pittsburgh, PA. HEPES and HEPES, sodium salt were products of BDH Chemicals, obtained through VWR International, West Chester, PA. CsCl was purchased from RPI, Arlington Heights, IL.

Inverted Microscopy:

Article Title: Probing Prokaryotic Social Behaviors with Bacterial "Lobster Traps"
Article Snippet: The beam was adjusted to overfill the back aperture of an oil immersion objective (Zeiss 100× Fluar, 1.3 numerical aperture [NA]) situated on a Zeiss Axiovert inverted microscope system. .. Structures were fabricated using 400 mg ml−1 BSA (Equitech-Bio) and 8.5 mM rose bengal (Sigma) as a photosensitizer in 20 mM HEPES–0.1 M NaCl buffer (pH 7.4) on no. 1 coverglass (VWR) within a single-pass flow cell system ( ).

Sonication:

Article Title: A Maltose-Binding Protein Fusion Construct Yields a Robust Crystallography Platform for MCL1
Article Snippet: .. Protein Purification of MCL1, MBP-MCL1 Fusion, TRX-MCL1 Fusion and Mutants Cells were resuspended in 50mM HEPES pH 7.5, 500mM NaCl, 5% glycerol, 0.001% 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) (JT Baker), 0.5mM Tris-(2-carboxyethyl)phosphine (TCEP) (VWR), 250 U Benzonase (Novagen), 100 mg lysozyme (Sigma) and one complete EDTA free protease inhibitor tablet (Roche), lysed via sonication and clarified via centrifugation. ..

Recombinant:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Recombinant gH-V5/gL-Flag complexes were precipitated from the lysates of 293T cells transfected with the respective expression constructs. .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

Molecular Weight:

Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug
Article Snippet: .. The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)). ..

Isolation:

Article Title: SAP97-mediated ADAM10 trafficking from Golgi outposts depends on PKC phosphorylation
Article Snippet: TIF and P3 preparation TIF and P3 were isolated from rat acute hippocampal slices. .. Slices were homogenized at 4 °C in an ice-cold lysis buffer with protease inhibitors, phosphatase inhibitors, 0.32 M sucrose, 1 mM HEPES, 0.1 mM PMSF, 1 mM MgCl2 using a glass-Teflon homogenizer (VWR, Radnor, PA, USA).

Transfection:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Recombinant gH-V5/gL-Flag complexes were precipitated from the lysates of 293T cells transfected with the respective expression constructs. .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

Purification:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: Paragraph title: Functional Reconstitution of Purified GLUT1 into Proteoliposomes ... 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C.

Protein Purification:

Article Title: A Maltose-Binding Protein Fusion Construct Yields a Robust Crystallography Platform for MCL1
Article Snippet: .. Protein Purification of MCL1, MBP-MCL1 Fusion, TRX-MCL1 Fusion and Mutants Cells were resuspended in 50mM HEPES pH 7.5, 500mM NaCl, 5% glycerol, 0.001% 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) (JT Baker), 0.5mM Tris-(2-carboxyethyl)phosphine (TCEP) (VWR), 250 U Benzonase (Novagen), 100 mg lysozyme (Sigma) and one complete EDTA free protease inhibitor tablet (Roche), lysed via sonication and clarified via centrifugation. ..

Selection:

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

Staining:

Article Title: Engineering triiodothyronine (T3) nanoparticle for use in ischemic brain stroke
Article Snippet: Coumarin-6 and 2,3,5-triphenyltetrazolium chloride (TTC) stain were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). .. HEPES, NaCl, and EDTA were received from VWR International (West Chester, PA, USA).

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

IA:

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

SDS Page:

Article Title: Potential adverse effects of botanical supplementation in high-fat-fed female mice
Article Snippet: Analysis of protein expression Skeletal muscle and liver lysates were prepared from powdered tissue by homogenizing in 25 mM HEPES, pH 7.4, 1% Igepal CA630, 137 mM NaCl, 1 mM PMSF, 10 μg/ml aprotinin, 1 μg/ml pepstatin, 5 μg/ml leupeptin, 10 mM Na4 P2 O7 , 100 mM NaF, and 2 mM NaVO4 using a Sonifier 450 homogenizer (VWR, Radnor, PA). .. The tissue supernatants (50 μg) were resolved by SDS-PAGE and subjected to immunoblotting using chemiluminescence detection (Thermo Fisher Scientific, Rockford, IL) and quantified as described [ ].

Article Title: Phosphatidic acid mediates the targeting of tBid to induce lysosomal membrane permeabilization and apoptosis [S]
Article Snippet: Experiments were performed in a Superdex-200 (1.5 × 45 cm) column equilibrated with 100 mM KCl, 10 mM HEPES, and 0.2 mM EDTA (pH 7.0) with or without 2% (w/v) CHAPS (J. T. Baker Inc.) at a 1 ml/min flow rate. .. Aliquots of individual fractions were subjected to SDS-PAGE in 15% Tris-glycine gels, followed by visualization of tBid using anti-Bid antibody.

Plasmid Preparation:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. After one wash, pre-coupled complexes were incubated overnight at 4°C with agitation with equal amounts of lysate of full-length EphA2-myc or full-length EphB3-myc expression plasmid transfected 293T cells (for KSHV gH/gL or RRV gH/gL binding analysis, respectively) or with supernatant of 293T cells transfected with an expression plasmid for HA-tagged EphA2 ectodomain (for KSHV gH/gL binding analysis).

SYBR Green Assay:

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

Functional Assay:

Article Title: Purification and Characterization of Mammalian Glucose Transporters Expressed in Pichia Pastoris
Article Snippet: Paragraph title: Functional Reconstitution of Purified GLUT1 into Proteoliposomes ... 12.5 mg of soybean phospholipids (Avanti Polar Lipids, Alabaster, AL) in 20 mM HEPES, 0.30 M NaCl were dispersed at 22°C in a bath sonicator (VWR M75D, VWR Int, Batavia, IL) and then mixed with 37 mM CHAPS and incubated for 2 hours at 25 °C.

Binding Assay:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. After one wash, pre-coupled complexes were incubated overnight at 4°C with agitation with equal amounts of lysate of full-length EphA2-myc or full-length EphB3-myc expression plasmid transfected 293T cells (for KSHV gH/gL or RRV gH/gL binding analysis, respectively) or with supernatant of 293T cells transfected with an expression plasmid for HA-tagged EphA2 ectodomain (for KSHV gH/gL binding analysis).

Size-exclusion Chromatography:

Article Title: Phosphatidic acid mediates the targeting of tBid to induce lysosomal membrane permeabilization and apoptosis [S]
Article Snippet: The oligomerization of tBid was measured by size-exclusion chromatography. .. Experiments were performed in a Superdex-200 (1.5 × 45 cm) column equilibrated with 100 mM KCl, 10 mM HEPES, and 0.2 mM EDTA (pH 7.0) with or without 2% (w/v) CHAPS (J. T. Baker Inc.) at a 1 ml/min flow rate.

Immunoprecipitation:

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: Paragraph title: Immunoprecipitation and Western blot analysis ... Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C.

Lysis:

Article Title: SAP97-mediated ADAM10 trafficking from Golgi outposts depends on PKC phosphorylation
Article Snippet: .. Slices were homogenized at 4 °C in an ice-cold lysis buffer with protease inhibitors, phosphatase inhibitors, 0.32 M sucrose, 1 mM HEPES, 0.1 mM PMSF, 1 mM MgCl2 using a glass-Teflon homogenizer (VWR, Radnor, PA, USA). ..

Article Title: Combination of Cannabinoids, Δ9- Tetrahydrocannabinol and Cannabidiol, Ameliorates Experimental Multiple Sclerosis by Suppressing Neuroinflammation Through Regulation of miRNA-Mediated Signaling Pathways
Article Snippet: .. Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). .. EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).

Article Title: A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus
Article Snippet: .. Lysates were prepared in NP40 lysis buffer (1% Nonidet P40 Substitute (Sigma-Aldrich), 150mM NaCl (Sigma-Aldrich), 50mM HEPES (VWR), 1mM EDTA (Amresco) with freshly added Protease Inhibitor Cocktail, General Use (Amresco)) and subsequently incubated with agitation with 0.5μg V5-tag antibody (Serotec or Bio-Rad) and ProteinG sepharose (GenScript or GE Healthcare) for 2h or overnight at 4°C. .. Amount of input gH/gL between mutants was normalized by diluting lysates with cell lysate from non-transfected 293T cells prior to immunoprecipitation according to Western blot analysis of lysates and evaluation of the gH/gL content for each construct.

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    Avantor hepes
    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml <t>ciprofloxacin</t> in <t>HEPES</t> buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.
    Hepes, supplied by Avantor, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze-thaw, blue diamonds), CFI containing 0.05% polysorbate 20 after freeze-thaw (red squares), CFI containing 0.1% polysorbate 20 after freeze-thaw (green triangles), and CFI containing 0.2% polysorbate 20 after freeze-thaw (yellow circles). Duplicate samples were analyzed at each time point.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation

    Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw in liquid nitrogen on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and 0, 0.05 or 0.1% polysorbate 20 after freeze-thaw. The CFI formulations (at 12.5 mg/ml) were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control prior to freeze-thaw ( blue diamonds ) or after freeze-thaw ( red circles ), CFI containing 0.05% polysorbate 20 after freeze-thaw ( green diamonds ), CFI containing 0.1% polysorbate 20 after freeze-thaw ( yellow triangles ). Duplicate samples were analyzed at each time point. Two simulated curves were generated by adjusting the release from the CFI sample without polysorbate 20 after freeze-thaw by assuming the addition of either 10% free drug ( green dashed line ) or 20% free drug ( yellow dashed line ) and normalizing the total drug release to 100%.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation, Generated

    Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.

    Journal: Pharmaceutical Research

    Article Title: Tuning Ciprofloxacin Release Profiles from Liposomally Encapsulated Nanocrystalline Drug

    doi: 10.1007/s11095-016-2002-5

    Figure Lengend Snippet: Evaluation of the effect of freeze-thaw at −50°C on the IVR profiles of CFI formulations at pH 6.0 containing 90 mg/ml sucrose and Brij 30 before ( a ) and after ( b ) freeze-thaw. The 12.5 mg/ml CFI formulations were diluted to 50 μg/ml ciprofloxacin in HEPES buffered saline (HBS) prior to a 1:1 dilution in bovine serum to measure the release of ciprofloxacin after incubation at 37°C for up to 4 h. IVR profiles are shown for the CFI control (no freeze thaw, blue diamonds ), CFI containing 0.05% Brij 30 ( red squares ), CFI containing 0.1% Brij 30 ( green triangles ), CFI containing 0.2% Brij 30 ( orange circles ), and CFI containing 0.3% Brij 30 ( purple squares ). Duplicate samples were analyzed at each time point.

    Article Snippet: The following materials were used for the preparation, characterization or analysis of the liposomal ciprofloxacin (CFI) formulations: sucrose (Sigma-Aldrich (St. Louis, MO)), HEPES, free acid (Avantor (Center Valley, PA)), sodium chloride (Amresco (Solon, OH)), HPLC grade methanol (Fisher Scientific (Fair Lawn, NJ)), triethylamine (TEA, JT Baker (USA)), polysorbate 20 (VWR Int. (West Chester, NJ)), polyethylene glycol dodecyl ether (Brij 30 (Sigma-Aldrich (Australia))), Donor Adult Bovine Serum (HyClone (Logan, Utah)), and Nanosep centrifugal filtration devices, 10 K and 30 K molecular weight (Pall Corporation (Ann Arbor, MI)).

    Techniques: Incubation