hepes buffer  (Millipore)


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    Structured Review

    Millipore hepes buffer
    Activity of the lysins against log-phase and stationary P. aeruginosa bacteria. The P. aeruginosa bacteria were grown overnight (stationary [Stat]), diluted 1:100, and grown to log phase (Log). Bacteria were washed and incubated with lysins at the indicated concentrations in <t>30 mM</t> <t>HEPES</t> buffer, pH 7.4, for 1 h at 37°C. Viable bacteria were quantified by serial dilution and plating. Experiments were done in duplicate; error bars represent standard deviations.
    Hepes Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 207 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hepes buffer/product/Millipore
    Average 99 stars, based on 207 article reviews
    Price from $9.99 to $1999.99
    hepes buffer - by Bioz Stars, 2020-03
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    Images

    1) Product Images from "Isolation of Phage Lysins That Effectively Kill Pseudomonas aeruginosa in Mouse Models of Lung and Skin Infection"

    Article Title: Isolation of Phage Lysins That Effectively Kill Pseudomonas aeruginosa in Mouse Models of Lung and Skin Infection

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00024-19

    Activity of the lysins against log-phase and stationary P. aeruginosa bacteria. The P. aeruginosa bacteria were grown overnight (stationary [Stat]), diluted 1:100, and grown to log phase (Log). Bacteria were washed and incubated with lysins at the indicated concentrations in 30 mM HEPES buffer, pH 7.4, for 1 h at 37°C. Viable bacteria were quantified by serial dilution and plating. Experiments were done in duplicate; error bars represent standard deviations.
    Figure Legend Snippet: Activity of the lysins against log-phase and stationary P. aeruginosa bacteria. The P. aeruginosa bacteria were grown overnight (stationary [Stat]), diluted 1:100, and grown to log phase (Log). Bacteria were washed and incubated with lysins at the indicated concentrations in 30 mM HEPES buffer, pH 7.4, for 1 h at 37°C. Viable bacteria were quantified by serial dilution and plating. Experiments were done in duplicate; error bars represent standard deviations.

    Techniques Used: Activity Assay, Incubation, Serial Dilution

    Activity of leading lysins against various bacteria. Various isolates of P. aeruginosa (A), Klebsiella and Enterobacter (B), and other Gram-negative and Gram-positive bacteria (C) were incubated with 100-μg/ml lysins in 30 mM HEPES buffer, pH 7.4, for 1 h at 37°C. Viable bacteria were enumerated by serial dilution and plating. Experiments were done in duplicate; error bars represent standard deviations. Also see Fig. S7 in the supplemental material.
    Figure Legend Snippet: Activity of leading lysins against various bacteria. Various isolates of P. aeruginosa (A), Klebsiella and Enterobacter (B), and other Gram-negative and Gram-positive bacteria (C) were incubated with 100-μg/ml lysins in 30 mM HEPES buffer, pH 7.4, for 1 h at 37°C. Viable bacteria were enumerated by serial dilution and plating. Experiments were done in duplicate; error bars represent standard deviations. Also see Fig. S7 in the supplemental material.

    Techniques Used: Activity Assay, Incubation, Serial Dilution

    Characterization of PlyPa03 and PlyPa91. (A) Time-kill curve. Log-phase P. aeruginosa PAO1 cells were incubated for various lengths of time at 37°C with 100-μg/ml lysin in 30 mM HEPES buffer. (B) Effect of pH. Log-phase P. aeruginosa PAO1 cells were incubated for 1 h at 37°C with 100-μg/ml lysin in 25 mM the following buffers: pH 5.0, acetate buffer; pH 6.0, MES buffer; pH 7.0 and 8.0, HEPES buffer; pH 9.0, CHES buffer; pH 10.0, CAPS buffer. (C) Effect of NaCl. Log-phase P. aeruginosa PAO1 cells were incubated with 100-μg/ml PlyPa03 or PlyPa91 for 1 h at 37°C in 30 mM HEPES, pH 7.4, and various concentrations of NaCl. (D) Effect of urea. Log-phase P. aeruginosa PAO1 cells were incubated with 100-μg/ml PlyPa03 or PlyPa91 for 1 h at 37°C in 30 mM HEPES, pH 7.4, and various concentrations of urea. In all cases, the surviving bacteria were enumerated by serial dilution and plating. Experiments were done in triplicate; error bars represent standard deviations.
    Figure Legend Snippet: Characterization of PlyPa03 and PlyPa91. (A) Time-kill curve. Log-phase P. aeruginosa PAO1 cells were incubated for various lengths of time at 37°C with 100-μg/ml lysin in 30 mM HEPES buffer. (B) Effect of pH. Log-phase P. aeruginosa PAO1 cells were incubated for 1 h at 37°C with 100-μg/ml lysin in 25 mM the following buffers: pH 5.0, acetate buffer; pH 6.0, MES buffer; pH 7.0 and 8.0, HEPES buffer; pH 9.0, CHES buffer; pH 10.0, CAPS buffer. (C) Effect of NaCl. Log-phase P. aeruginosa PAO1 cells were incubated with 100-μg/ml PlyPa03 or PlyPa91 for 1 h at 37°C in 30 mM HEPES, pH 7.4, and various concentrations of NaCl. (D) Effect of urea. Log-phase P. aeruginosa PAO1 cells were incubated with 100-μg/ml PlyPa03 or PlyPa91 for 1 h at 37°C in 30 mM HEPES, pH 7.4, and various concentrations of urea. In all cases, the surviving bacteria were enumerated by serial dilution and plating. Experiments were done in triplicate; error bars represent standard deviations.

    Techniques Used: Incubation, Serial Dilution

    Bactericidal activity of lysins against P. aeruginosa PAO1. Purified lysins were diluted to various concentrations and incubated with log-phase P. aeruginosa PAO1 for 1 h at 37°C in 30 mM HEPES, pH 7.4. The values of the number of CFU per milliliter were established by serial dilution and plating. (A) Initial lysins. (B) Additional homologues of PlyPa02. Experiments were conducted in duplicate; error bars represent standard deviations.
    Figure Legend Snippet: Bactericidal activity of lysins against P. aeruginosa PAO1. Purified lysins were diluted to various concentrations and incubated with log-phase P. aeruginosa PAO1 for 1 h at 37°C in 30 mM HEPES, pH 7.4. The values of the number of CFU per milliliter were established by serial dilution and plating. (A) Initial lysins. (B) Additional homologues of PlyPa02. Experiments were conducted in duplicate; error bars represent standard deviations.

    Techniques Used: Activity Assay, Purification, Incubation, Serial Dilution

    Related Articles

    Centrifugation:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: After centrifugation, tryptase concentration in the supernatant was quantified using a Mouse Mast Cell Tryptase (MCT) ELISA Kit (Cusabio). .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate.

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Bacterial cells were lysed by sonication and cell debris was removed by centrifugation. .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Positive Control:

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma). .. Spleen cells were cultured with medium alone, the optimal concentration of OVA (5.0 g/mL) or PHA or PMA + anti-mouse CD3 (BD Biosciences) as positive control stimuli.

    Synthesized:

    Article Title: Comparison of Copper(II)-Ligand Complexes as Mediators for Preparing Electrochemically Modulated Nitric Oxide-Releasing Catheters
    Article Snippet: 1,4,7-Trimethyl-1,4,7-triazacyclononane (Me3 TACN), tris(2-pyridylmethyl)amine (TPMA), copper(II) sulfate pentahydrate, disodium phosphate, HEPES buffer, potassium chloride, potassium phosphate monobasic, sodium chloride, sodium nitrite, and sodium phosphate were obtained from Sigma-Aldrich (St. Louis, MO). .. N-propanoate-N,N-bis(2-pyridylmethyl)amine (BMPA-Pr) and N-propanoate-N,N-bis(2-pyridylethyl)amine (BEPA-Pr) were synthesized in accordance with the procedures included in the .

    Autoradiography:

    Article Title: Marked decrease of neuropeptide Y Y2 receptor binding sites in the hippocampus in murine prion disease
    Article Snippet: After air-dry, the sections were incubated in Hepes buffer (pH 7.4) for 60 min at room temperature, followed by preincubation for 20 min at room temperature in Hepes buffer containing 0.1% BSA (Sigma) and 0.05% Bacitracin (Sigma), in some cases with NPY (porcine; Bachem) or the Y2 agonist NPY ( – ) (porcine; Peninsula Laboratories) in Hepes buffer with BSA and Bacitracin. .. The slides were exposed to an autoradiography film (Hyperfilm-3 H, Amersham) for 3–6 days at −20°C, developed for 5 min in LX 24 (Kodak), fixed in AL4 (Kodak) for 13 min, and finally rinsed in running water and air dried.

    Live Dead Assay:

    Article Title: Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering
    Article Snippet: Cell viability was assessed by Live/Dead assay (Invitrogen) immediately after encapsulation to verify that the encapsulation process was cytocompatible ( ). .. Chondrocyte-hydrogel constructs were cultured in chondrocyte growth medium composed of high-glucose DMEM (Gibco) containing 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin and fungizone (Gibco, Invitrogen), 50 mg/mL L-ascorbate-2-phosphate (Sigma-Aldrich), 40 mg/mL L-proline (Sigma-Aldrich), 100 mg/mL non-essential amino acids (Gibco), 100 mg/mL HEPES buffer (Sigma-Aldrich) and 50 mg/mL gentamicin (Invitrogen).

    Construct:

    Article Title: Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering
    Article Snippet: .. Chondrocyte-hydrogel constructs were cultured in chondrocyte growth medium composed of high-glucose DMEM (Gibco) containing 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin and fungizone (Gibco, Invitrogen), 50 mg/mL L-ascorbate-2-phosphate (Sigma-Aldrich), 40 mg/mL L-proline (Sigma-Aldrich), 100 mg/mL non-essential amino acids (Gibco), 100 mg/mL HEPES buffer (Sigma-Aldrich) and 50 mg/mL gentamicin (Invitrogen). .. Medium was changed every other day, and cell-hydrogel constructs were maintained with 5% CO2 at 37°C.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: After centrifugation, tryptase concentration in the supernatant was quantified using a Mouse Mast Cell Tryptase (MCT) ELISA Kit (Cusabio). .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate.

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma). .. Supernatants were collected after 72 h and assayed for mouse IFN-␥, IL-4, IL-5 and IL-10 using ELISA assays (Quantikine brand, R Systems, Minneapolis, MN) according to the manufacturer's protocol.

    Incubation:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate. .. After 90-min incubation at 37°C, the reaction was stopped with glycine buffer (400 mM, pH 10.7).

    Article Title: Isolation of Phage Lysins That Effectively Kill Pseudomonas aeruginosa in Mouse Models of Lung and Skin Infection
    Article Snippet: For time-kill curves, following incubation, assay contents were diluted 1:1 in 5% BBL beef extract (BD) to stop the reaction and were immediately diluted and plated. .. Evaluation of the effect of serum was done in 30 mM HEPES buffer, pH 7.4, 100-μg/ml lysins, using serially diluted pooled human type AB blood serum from male subjects (Sigma).

    Article Title: Common pathways for receptor-mediated ingestion of Escherichia coli and LDL cholesterol by Entamoeba histolytica regulated in part by transmembrane kinase 39
    Article Snippet: .. Cells (2 × 108 ) were washed once in HEPES buffer (10 mM HEPES, 140 nM sodium chloride, pH 7.2) supplemented with 0.1% heat-inactivated BSA and resuspended to a concentration of 1 × 108 cells/ml in HEPES buffer with or without calcium (2.5 mM calcium chloride) and incubated at 37°C for 48 h. After 48 h erythrocytes were stained with PKH26 Red Fluorescent Cell Linker dye (PKH26-GL; Sigma Aldrich, St. Louis, MO, USA) according to the manufacturer’s protocol. .. Briefly, erythrocytes were harvested by centrifugation, washed once in 0.1% BSA-HEPES buffer, resuspended in 1 ml of Diluent C (kit component proprietary from Sigma Aldrich), and added to 1 ml of 4 μM PKH26 dye in Diluent C. Following incubation at 25°C for 5 min, stained erythrocytes were added to 2 ml of 1% BSA-HEPES buffer and washed three times in M199s medium (5.7 mM D-cysteine and 25 mM HEPES, pH 7.2) + 0.5% BSA.

    Article Title: Marked decrease of neuropeptide Y Y2 receptor binding sites in the hippocampus in murine prion disease
    Article Snippet: .. After air-dry, the sections were incubated in Hepes buffer (pH 7.4) for 60 min at room temperature, followed by preincubation for 20 min at room temperature in Hepes buffer containing 0.1% BSA (Sigma) and 0.05% Bacitracin (Sigma), in some cases with NPY (porcine; Bachem) or the Y2 agonist NPY ( – ) (porcine; Peninsula Laboratories) in Hepes buffer with BSA and Bacitracin. .. After rinsing, sections were incubated in monoiodinated peptide YY (125 I-PYY) (0.1 nM) (DuPont/NEN) or in 125 I-PYY plus 10−6 M of either NPY ligand for 60 min. Then, the sections were rinsed in ice-cold buffer, dried by a cold air stream, and stored in a dessicator over night.

    Activity Assay:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate. .. After 90-min incubation at 37°C, the reaction was stopped with glycine buffer (400 mM, pH 10.7).

    Infection:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma). .. After removing cell debris by centrifugation, the virus-infected cell lysate was heated for 30 min at 55°C to inactivate any live virus.

    Expressing:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Expression of the N protein was induced by adding 0.1 mM IPTG for 6 h at 25°C in Escherichia coli strain B834. .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Modification:

    Article Title: Novel and potent anti-tumor and anti-metastatic di-2-pyridylketone thiosemicarbazones demonstrate marked differences in pharmacology between the first and second generation lead agents
    Article Snippet: .. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Lonza, Switzerland) with or without (for MCF-7 cells only) phenol red, supplemented with 10% heat-inactivated fetal bovine serum (FBS; Lonza, Switzerland), 1% penicillin/streptomycin solution (Lonza, Switzerland) and 10 mM HEPES buffer (pH 7.0-7.6; Sigma-Aldrich, Germany). .. The HL-60 cell line was maintained in RPMI medium (Sigma-Aldrich, Germany) supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin solution.

    Colorimetric Assay:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate. .. After 90-min incubation at 37°C, the reaction was stopped with glycine buffer (400 mM, pH 10.7).

    Chromatography:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Nucleocapsid proteins in soluble fraction were purified using a combination of Ni-NTA agarose chromatography (Qiagen) and S-200 size exclusion chromatography (Amersham Biosciences). .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Concentration Assay:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: After centrifugation, tryptase concentration in the supernatant was quantified using a Mouse Mast Cell Tryptase (MCT) ELISA Kit (Cusabio). .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate.

    Article Title: Isolation of Phage Lysins That Effectively Kill Pseudomonas aeruginosa in Mouse Models of Lung and Skin Infection
    Article Snippet: Assays evaluating the effect of pH were done by adding 25 μl of 100 mM the following buffers to wells of a 96-well plate (final concentration, 25 mM): pH 5.0, acetate buffer; pH 6.0, MES (morpholineethanesulfonic acid) buffer; pH 7.0 and 8.0, HEPES buffer; pH 9.0, N -cyclohexyl-2-aminoethanesulfonic acid (CHES) buffer; pH 10.0, N -cyclohexyl-3-aminopropanesulfonic acid (CAPS) buffer. .. Evaluation of the effect of serum was done in 30 mM HEPES buffer, pH 7.4, 100-μg/ml lysins, using serially diluted pooled human type AB blood serum from male subjects (Sigma).

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma). .. Spleen cells were cultured with medium alone, the optimal concentration of OVA (5.0 g/mL) or PHA or PMA + anti-mouse CD3 (BD Biosciences) as positive control stimuli.

    Article Title: Common pathways for receptor-mediated ingestion of Escherichia coli and LDL cholesterol by Entamoeba histolytica regulated in part by transmembrane kinase 39
    Article Snippet: .. Cells (2 × 108 ) were washed once in HEPES buffer (10 mM HEPES, 140 nM sodium chloride, pH 7.2) supplemented with 0.1% heat-inactivated BSA and resuspended to a concentration of 1 × 108 cells/ml in HEPES buffer with or without calcium (2.5 mM calcium chloride) and incubated at 37°C for 48 h. After 48 h erythrocytes were stained with PKH26 Red Fluorescent Cell Linker dye (PKH26-GL; Sigma Aldrich, St. Louis, MO, USA) according to the manufacturer’s protocol. .. Briefly, erythrocytes were harvested by centrifugation, washed once in 0.1% BSA-HEPES buffer, resuspended in 1 ml of Diluent C (kit component proprietary from Sigma Aldrich), and added to 1 ml of 4 μM PKH26 dye in Diluent C. Following incubation at 25°C for 5 min, stained erythrocytes were added to 2 ml of 1% BSA-HEPES buffer and washed three times in M199s medium (5.7 mM D-cysteine and 25 mM HEPES, pH 7.2) + 0.5% BSA.

    Protease Inhibitor:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma). .. After removing cell debris by centrifugation, the virus-infected cell lysate was heated for 30 min at 55°C to inactivate any live virus.

    Cell Culture:

    Article Title: Non-plaque-forming virions of Modified Vaccinia virus Ankara express viral genes.
    Article Snippet: .. Cells DF-1, HaCat and MA-104 Jena cells were cultured in VLE Dulbecco's MEM (Biochrom GmbH) supplemented with 10% heat inactivated fetal bovine serum (HI-FBS) (SIGMA), 1% Penicillin-Streptomycin (SIGMA) and 2% Hepes-buffer (SIGMA). .. HeLa cells were maintained in Minimum Essential Medium Eagle (MEM) (SIGMA) containing 10% HI-FBS and antibiotics as described above.

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Vero cells (ATCC CRL-1586) were cultured in DMEM supplemented with 5% fetal calf serum (Gibco) at 37°C with 5% CO 2 in a humidified incubator. .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma). .. Cells were counted for viability (always ≥95%) and cultured in RPMI + 10% FBS in quadruplicate wells at 4 × 10 5 per well in round-bottomed 96-well culture plates (Nunc brand, Fisher Scientific) for 72 h at 37 • C in a humidified 5% CO 2 incubator.

    Article Title: Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering
    Article Snippet: .. Chondrocyte-hydrogel constructs were cultured in chondrocyte growth medium composed of high-glucose DMEM (Gibco) containing 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin and fungizone (Gibco, Invitrogen), 50 mg/mL L-ascorbate-2-phosphate (Sigma-Aldrich), 40 mg/mL L-proline (Sigma-Aldrich), 100 mg/mL non-essential amino acids (Gibco), 100 mg/mL HEPES buffer (Sigma-Aldrich) and 50 mg/mL gentamicin (Invitrogen). .. Medium was changed every other day, and cell-hydrogel constructs were maintained with 5% CO2 at 37°C.

    Article Title: Novel and potent anti-tumor and anti-metastatic di-2-pyridylketone thiosemicarbazones demonstrate marked differences in pharmacology between the first and second generation lead agents
    Article Snippet: .. Cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Lonza, Switzerland) with or without (for MCF-7 cells only) phenol red, supplemented with 10% heat-inactivated fetal bovine serum (FBS; Lonza, Switzerland), 1% penicillin/streptomycin solution (Lonza, Switzerland) and 10 mM HEPES buffer (pH 7.0-7.6; Sigma-Aldrich, Germany). .. The HL-60 cell line was maintained in RPMI medium (Sigma-Aldrich, Germany) supplemented with 10% heat-inactivated FBS and 1% penicillin/streptomycin solution.

    Generated:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: A standard curve was generated using a four-parameter logistic (4-PL) curve fit. .. Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate.

    Sonication:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Bacterial cells were lysed by sonication and cell debris was removed by centrifugation. .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Binding Assay:

    Article Title: Marked decrease of neuropeptide Y Y2 receptor binding sites in the hippocampus in murine prion disease
    Article Snippet: Paragraph title: Autoradiographic Binding. ... After air-dry, the sections were incubated in Hepes buffer (pH 7.4) for 60 min at room temperature, followed by preincubation for 20 min at room temperature in Hepes buffer containing 0.1% BSA (Sigma) and 0.05% Bacitracin (Sigma), in some cases with NPY (porcine; Bachem) or the Y2 agonist NPY ( – ) (porcine; Peninsula Laboratories) in Hepes buffer with BSA and Bacitracin.

    Molecular Weight:

    Article Title: The role of the amorphous phase on the biomimetic mineralization of collagen
    Article Snippet: .. Collagen mineralization in presence of polyaspartic acid (pAsp) was achieved by incubating the collagen-adsorbed cryoTEM grids in Hepes buffer (10 mH, pH 7.4, Sigma) containing CaCl2 (2.7 mM, Merck), K2 HPO4 (1.35 mM, Merck) and polyaspartic acid (10 μg/ml, molecular weight 2000-11000 Da, Sigma) at 37 °C, as described elsewhere., Collagen mineralization in presence of C-DMP1 was achieved by incubating collagen-adsorbed cryoTEM grids in Hepes buffer (10 mM, pH 7.4, Sigma) containing CaCl2 (2.7 mM, Merck), K2 HPO4 (1.35 mM, Merck) and C-DMP1 (15 μg/ml, see below) + polyaspartic acid (1.5 μg/ml – 10 μg/ml, molecular weight 2000-11000 Da, Sigma) at 37 °C. ..

    Cell Stimulation:

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Paragraph title: Spleen cell stimulation and measurement of cytokines ... Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma).

    Isolation:

    Article Title: Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering
    Article Snippet: Paragraph title: 2.5. Chondrocyte isolation, encapsulation and cell culture ... Chondrocyte-hydrogel constructs were cultured in chondrocyte growth medium composed of high-glucose DMEM (Gibco) containing 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin and fungizone (Gibco, Invitrogen), 50 mg/mL L-ascorbate-2-phosphate (Sigma-Aldrich), 40 mg/mL L-proline (Sigma-Aldrich), 100 mg/mL non-essential amino acids (Gibco), 100 mg/mL HEPES buffer (Sigma-Aldrich) and 50 mg/mL gentamicin (Invitrogen).

    Size-exclusion Chromatography:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Nucleocapsid proteins in soluble fraction were purified using a combination of Ni-NTA agarose chromatography (Qiagen) and S-200 size exclusion chromatography (Amersham Biosciences). .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Purification:

    Article Title: Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11.
    Article Snippet: Nucleocapsid proteins in soluble fraction were purified using a combination of Ni-NTA agarose chromatography (Qiagen) and S-200 size exclusion chromatography (Amersham Biosciences). .. The Vero cells were infected with a SARS-CoV strain (CUHK-W1) for 16-48 h, and the cells were lysed in a Hepes buffer (10 mM; pH 7.0) supplemented with 40 mM KCl, 3 mM MgCl 2 , 5% glycerol, 0.2% NP40, 1 mM DTT, 1 mM PMSF, and 1· protease inhibitor cocktail (Sigma).

    Mouse Assay:

    Article Title: rOv-ASP-1, a recombinant secreted protein of the helminth Onchocercavolvulus, is a potent adjuvant for inducing antibodies to ovalbumin, HIV-1 polypeptide and SARS-CoV peptide antigens.
    Article Snippet: Spleen cell stimulation and measurement of cytokines After exsanguination of the mice under anaesthesia, spleens were removed, cut into two pieces with sterile scissors and made into single cell suspensions using sterile glass Potter-Elvehjem Tissue Grinders (Fisher Scientific International, Pittsburgh, PA). .. Cells were washed three times in RPMI supplemented with 2% heatinactivated foetal bovine serum (FBS), 10 mM HEPES buffer, 0.2 mM l-glutamine, 50 m 2-mercaptoethanol, 100 U/mL penicillin and 100 g/mL streptomycin (all from Sigma).

    In Vitro:

    Article Title: Hydrazone covalent adaptable networks modulate extracellular matrix deposition for cartilage tissue engineering
    Article Snippet: [ ] Freshly isolated porcine chondrocytes were encapsulated within 40 μL hydrazone hydrogels formed in 1 mL syringe barrels at ~50 million cells/mL, as this density has been shown to produce high quality neotissue for in vitro encapsulation studies. .. Chondrocyte-hydrogel constructs were cultured in chondrocyte growth medium composed of high-glucose DMEM (Gibco) containing 10% fetal bovine serum (Gibco), 1% penicillin-streptomycin and fungizone (Gibco, Invitrogen), 50 mg/mL L-ascorbate-2-phosphate (Sigma-Aldrich), 40 mg/mL L-proline (Sigma-Aldrich), 100 mg/mL non-essential amino acids (Gibco), 100 mg/mL HEPES buffer (Sigma-Aldrich) and 50 mg/mL gentamicin (Invitrogen).

    Activation Assay:

    Article Title: Rapid Mast Cell Generation from Gata2 Reporter Pluripotent Stem Cells
    Article Snippet: Paragraph title: Mast Cell Activation/Degranulation Analyses ... Following three washes with HEPES buffer (pH 7.4), 50,000 cells/well were activated with DNP-HSA (Sigma-Aldrich) and incubated in 37°C for 60 min. Supernatants were collected and cells lysed using 0.1% Triton X-100 in HEPES buffer. β-Hexosaminidase activity in supernatants and cell lysates was measured using a colorimetric assay with 3.5 mg/mL p-nitrophenyl-N-acetyl-β-D-glucosamide (PNAG; Sigma-Aldrich) as a substrate.

    Staining:

    Article Title: Common pathways for receptor-mediated ingestion of Escherichia coli and LDL cholesterol by Entamoeba histolytica regulated in part by transmembrane kinase 39
    Article Snippet: .. Cells (2 × 108 ) were washed once in HEPES buffer (10 mM HEPES, 140 nM sodium chloride, pH 7.2) supplemented with 0.1% heat-inactivated BSA and resuspended to a concentration of 1 × 108 cells/ml in HEPES buffer with or without calcium (2.5 mM calcium chloride) and incubated at 37°C for 48 h. After 48 h erythrocytes were stained with PKH26 Red Fluorescent Cell Linker dye (PKH26-GL; Sigma Aldrich, St. Louis, MO, USA) according to the manufacturer’s protocol. .. Briefly, erythrocytes were harvested by centrifugation, washed once in 0.1% BSA-HEPES buffer, resuspended in 1 ml of Diluent C (kit component proprietary from Sigma Aldrich), and added to 1 ml of 4 μM PKH26 dye in Diluent C. Following incubation at 25°C for 5 min, stained erythrocytes were added to 2 ml of 1% BSA-HEPES buffer and washed three times in M199s medium (5.7 mM D-cysteine and 25 mM HEPES, pH 7.2) + 0.5% BSA.

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  • 79
    Millipore hepes buffered serum supplemented dmem
    Entry phase and dose response. (A) Cell cultures (2 × 10 5 cells/well) in 96-well strip plates were switched to <t>HEPES-buffered</t> serum-supplemented <t>DMEM,</t> cooled on ice for 30 min, and infected at 4°C with 6 × 10 3 PFU of hr R3 per well. Cells were kept at 4°C for an additional 1 h before they were warmed to 23°C for 30 min and 37°C for the rest of the experiment. At 1-h intervals immediately following infection, strips of wells were treated for 1-h periods with 50 μM EB in serum-free DMEM and returned to regular medium. Before and after each treatment, the cells were rinsed three times with serum-free DMEM and serum-supplemented DMEM, respectively. The number of lacZ + cells was scored 11 h postinfection (●) and normalized to the number counted in mock-treated controls (maximally 187 ± 20 [ n = 3]). Separate strips of infected control cells were fixed and stained immediately following each mock treatment to monitor β-galactosidase expression over time (○). Data points represent means of triplicate measurements with standard errors of the means. (B) Cell cultures (2 × 10 5 cells/well) were switched to serum-free DMEM, cooled on ice, and infected at 4°C with 2.4 × 10 3 PFU of hr R3 per well. After infection, the cells were rinsed and treated with EB (●) or EBX (○) for 1 h at 4°C and for additional 30-min periods at 23 and 37°C. Triplicate counts of lacZ + cells were performed 6 h later (all points are means and standard errors of means; control score, 141 ± 5.9).
    Hepes Buffered Serum Supplemented Dmem, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore hepes electrochemistry buffer
    <t>p58C</t> 266-464 participates in redox switching on DNA. The cartoon (top left) depicts p58C DNA binding and redox switching on an Au electrode. (Bottom left) CV scan of electrochemically unaltered p58C 266-464 . (Right) bulk oxidation (above) of p58C 266-464 and subsequent CV scans (below). This construct displays similar electrochemical behavior to p58C 272-464 . All electrochemistry was performed in anaerobic conditions with 40 μM [4Fe4S] p58C 266-464 in 20 mM <t>HEPES</t> (pH 7.2), 75 mM NaCl. CV was performed at 100 mV/s scan rate.
    Hepes Electrochemistry Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore hepes naoh buffer
    The effects of pH and temperature on the activities and stabilities of Teth514_1788 and Teth514_1789. ( A ) The thermal stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at a temperature range between 30–90°C for 30 min. ( B ) The pH stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at 4°C for 24 h. ( C ) The pH dependence on the phosphorolytic and synthetic activities of Teth514_1788 (44 n m ) in 40 m m sodium citrate (pH 3.0–5.5), bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane-HCl (pH 5.5–7.0), <t>HEPES-NaOH</t> (pH 7.0–8.5), and glycine-NaOH (pH 8.5–10.5). ( D ) The pH dependence of the phosphorolytic and synthetic activities of Teth514_1789 (32 n m ) in the same buffers listed in Panel C. In Panels C and D, the closed and open symbols represent the synthetic and phosphorolytic activities, respectively.
    Hepes Naoh Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Millipore divalent cation free hepes tyrode buffer
    Activation-independent platelet aggregation mediated by multimeric VWF or isolated VWF A1 domain. Perfusion over immobilized VWF of washed blood cells suspended in <t>Hepes/Tyrode</t> buffer, pH 7.4 (see legend to ). In the presence of soluble multimeric
    Divalent Cation Free Hepes Tyrode Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Entry phase and dose response. (A) Cell cultures (2 × 10 5 cells/well) in 96-well strip plates were switched to HEPES-buffered serum-supplemented DMEM, cooled on ice for 30 min, and infected at 4°C with 6 × 10 3 PFU of hr R3 per well. Cells were kept at 4°C for an additional 1 h before they were warmed to 23°C for 30 min and 37°C for the rest of the experiment. At 1-h intervals immediately following infection, strips of wells were treated for 1-h periods with 50 μM EB in serum-free DMEM and returned to regular medium. Before and after each treatment, the cells were rinsed three times with serum-free DMEM and serum-supplemented DMEM, respectively. The number of lacZ + cells was scored 11 h postinfection (●) and normalized to the number counted in mock-treated controls (maximally 187 ± 20 [ n = 3]). Separate strips of infected control cells were fixed and stained immediately following each mock treatment to monitor β-galactosidase expression over time (○). Data points represent means of triplicate measurements with standard errors of the means. (B) Cell cultures (2 × 10 5 cells/well) were switched to serum-free DMEM, cooled on ice, and infected at 4°C with 2.4 × 10 3 PFU of hr R3 per well. After infection, the cells were rinsed and treated with EB (●) or EBX (○) for 1 h at 4°C and for additional 30-min periods at 23 and 37°C. Triplicate counts of lacZ + cells were performed 6 h later (all points are means and standard errors of means; control score, 141 ± 5.9).

    Journal: Journal of Virology

    Article Title: Modified FGF4 Signal Peptide Inhibits Entry of Herpes Simplex Virus Type 1

    doi: 10.1128/JVI.75.6.2634-2645.2001

    Figure Lengend Snippet: Entry phase and dose response. (A) Cell cultures (2 × 10 5 cells/well) in 96-well strip plates were switched to HEPES-buffered serum-supplemented DMEM, cooled on ice for 30 min, and infected at 4°C with 6 × 10 3 PFU of hr R3 per well. Cells were kept at 4°C for an additional 1 h before they were warmed to 23°C for 30 min and 37°C for the rest of the experiment. At 1-h intervals immediately following infection, strips of wells were treated for 1-h periods with 50 μM EB in serum-free DMEM and returned to regular medium. Before and after each treatment, the cells were rinsed three times with serum-free DMEM and serum-supplemented DMEM, respectively. The number of lacZ + cells was scored 11 h postinfection (●) and normalized to the number counted in mock-treated controls (maximally 187 ± 20 [ n = 3]). Separate strips of infected control cells were fixed and stained immediately following each mock treatment to monitor β-galactosidase expression over time (○). Data points represent means of triplicate measurements with standard errors of the means. (B) Cell cultures (2 × 10 5 cells/well) were switched to serum-free DMEM, cooled on ice, and infected at 4°C with 2.4 × 10 3 PFU of hr R3 per well. After infection, the cells were rinsed and treated with EB (●) or EBX (○) for 1 h at 4°C and for additional 30-min periods at 23 and 37°C. Triplicate counts of lacZ + cells were performed 6 h later (all points are means and standard errors of means; control score, 141 ± 5.9).

    Article Snippet: In some experiments, aliquots of diluted virus were first dialyzed (Spectra/Por; molecular weight cutoff, 12,000 to 14,000) overnight at 4°C against a 60-fold excess volume of HEPES-buffered serum-supplemented DMEM or forced by syringe through 0.22-μm-pore-size membranes (Millex-GV; Millipore) before the remaining infectious virus was assayed.

    Techniques: Stripping Membranes, Infection, Staining, Expressing

    Effect of pretreating cells with EB on subsequent virus infection. Cell cultures (2 × 10 5 cells/well) in microtiter wells were switched to serum-free HEPES-buffered DMEM and incubated for 30 min at 37°C. A first set of cells were then pretreated for 1 h with EB, rinsed three times, and infected for 1 h with 7,400 PFU of hr R3 per well before they were returned to regular medium. Rinses and infections were carried out in the absence of EB in either serum-free (Δ) or serum-supplemented (▴) DMEM. A second set of cells was pretreated for 1 h with (●) or without (○) EB and infected for 1 h with 7,400 PFU of hr R3 per well in the presence of EB before they were returned to regular medium. Triplicate counts of lacZ + cells were performed 8 h postinfection (all points are means with standard errors of the means; control score, 265 ± 13 [ n = 3]).

    Journal: Journal of Virology

    Article Title: Modified FGF4 Signal Peptide Inhibits Entry of Herpes Simplex Virus Type 1

    doi: 10.1128/JVI.75.6.2634-2645.2001

    Figure Lengend Snippet: Effect of pretreating cells with EB on subsequent virus infection. Cell cultures (2 × 10 5 cells/well) in microtiter wells were switched to serum-free HEPES-buffered DMEM and incubated for 30 min at 37°C. A first set of cells were then pretreated for 1 h with EB, rinsed three times, and infected for 1 h with 7,400 PFU of hr R3 per well before they were returned to regular medium. Rinses and infections were carried out in the absence of EB in either serum-free (Δ) or serum-supplemented (▴) DMEM. A second set of cells was pretreated for 1 h with (●) or without (○) EB and infected for 1 h with 7,400 PFU of hr R3 per well in the presence of EB before they were returned to regular medium. Triplicate counts of lacZ + cells were performed 8 h postinfection (all points are means with standard errors of the means; control score, 265 ± 13 [ n = 3]).

    Article Snippet: In some experiments, aliquots of diluted virus were first dialyzed (Spectra/Por; molecular weight cutoff, 12,000 to 14,000) overnight at 4°C against a 60-fold excess volume of HEPES-buffered serum-supplemented DMEM or forced by syringe through 0.22-μm-pore-size membranes (Millex-GV; Millipore) before the remaining infectious virus was assayed.

    Techniques: Infection, Incubation

    p58C 266-464 participates in redox switching on DNA. The cartoon (top left) depicts p58C DNA binding and redox switching on an Au electrode. (Bottom left) CV scan of electrochemically unaltered p58C 266-464 . (Right) bulk oxidation (above) of p58C 266-464 and subsequent CV scans (below). This construct displays similar electrochemical behavior to p58C 272-464 . All electrochemistry was performed in anaerobic conditions with 40 μM [4Fe4S] p58C 266-464 in 20 mM HEPES (pH 7.2), 75 mM NaCl. CV was performed at 100 mV/s scan rate.

    Journal: PLoS ONE

    Article Title: Functional and structural similarity of human DNA primase [4Fe4S] cluster domain constructs

    doi: 10.1371/journal.pone.0209345

    Figure Lengend Snippet: p58C 266-464 participates in redox switching on DNA. The cartoon (top left) depicts p58C DNA binding and redox switching on an Au electrode. (Bottom left) CV scan of electrochemically unaltered p58C 266-464 . (Right) bulk oxidation (above) of p58C 266-464 and subsequent CV scans (below). This construct displays similar electrochemical behavior to p58C 272-464 . All electrochemistry was performed in anaerobic conditions with 40 μM [4Fe4S] p58C 266-464 in 20 mM HEPES (pH 7.2), 75 mM NaCl. CV was performed at 100 mV/s scan rate.

    Article Snippet: All p58C constructs were buffer exchanged into HEPES electrochemistry buffer (20 mM HEPES (pH 7.2), 75 mM NaCl) using Amicon ultra centrifugal filters (0.5 mL, 3 kDa MWCO) (Millipore Sigma).

    Techniques: Binding Assay, Construct

    The effects of pH and temperature on the activities and stabilities of Teth514_1788 and Teth514_1789. ( A ) The thermal stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at a temperature range between 30–90°C for 30 min. ( B ) The pH stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at 4°C for 24 h. ( C ) The pH dependence on the phosphorolytic and synthetic activities of Teth514_1788 (44 n m ) in 40 m m sodium citrate (pH 3.0–5.5), bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane-HCl (pH 5.5–7.0), HEPES-NaOH (pH 7.0–8.5), and glycine-NaOH (pH 8.5–10.5). ( D ) The pH dependence of the phosphorolytic and synthetic activities of Teth514_1789 (32 n m ) in the same buffers listed in Panel C. In Panels C and D, the closed and open symbols represent the synthetic and phosphorolytic activities, respectively.

    Journal: PLoS ONE

    Article Title: Discovery of Two β-1,2-Mannoside Phosphorylases Showing Different Chain-Length Specificities from Thermoanaerobacter sp. X-514

    doi: 10.1371/journal.pone.0114882

    Figure Lengend Snippet: The effects of pH and temperature on the activities and stabilities of Teth514_1788 and Teth514_1789. ( A ) The thermal stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at a temperature range between 30–90°C for 30 min. ( B ) The pH stabilities of 360 n m Teth514_1788 (closed symbols) and 320 n m Teth514_1789 (open symbols) at 4°C for 24 h. ( C ) The pH dependence on the phosphorolytic and synthetic activities of Teth514_1788 (44 n m ) in 40 m m sodium citrate (pH 3.0–5.5), bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane-HCl (pH 5.5–7.0), HEPES-NaOH (pH 7.0–8.5), and glycine-NaOH (pH 8.5–10.5). ( D ) The pH dependence of the phosphorolytic and synthetic activities of Teth514_1789 (32 n m ) in the same buffers listed in Panel C. In Panels C and D, the closed and open symbols represent the synthetic and phosphorolytic activities, respectively.

    Article Snippet: After washing with buffer A containing 22 mm imidazole and subsequently eluting the proteins with a 22–400 mm imidazole linear gradient in buffer A, the fractions containing the recombinant protein were pooled, dialyzed against 10 mm HEPES-NaOH buffer (pH 7.0), and concentrated (AMICON Ultra-15 filter; Millipore, Billerica, MA, USA).

    Techniques:

    Activation-independent platelet aggregation mediated by multimeric VWF or isolated VWF A1 domain. Perfusion over immobilized VWF of washed blood cells suspended in Hepes/Tyrode buffer, pH 7.4 (see legend to ). In the presence of soluble multimeric

    Journal:

    Article Title: Activation-independent platelet adhesion and aggregation under elevated shear stress

    doi: 10.1182/blood-2006-04-011551

    Figure Lengend Snippet: Activation-independent platelet aggregation mediated by multimeric VWF or isolated VWF A1 domain. Perfusion over immobilized VWF of washed blood cells suspended in Hepes/Tyrode buffer, pH 7.4 (see legend to ). In the presence of soluble multimeric

    Article Snippet: This procedure was repeated twice, each time using half the amount of apyrase, and after the final centrifugation the cells were suspended in divalent cation-free Hepes/Tyrode buffer, pH 7.4, containing 50 mg/mL bovine serum albumin (BSA; Calbiochem, La Jolla, CA).

    Techniques: Activation Assay, Isolation