phosphate buffered saline pbs without cacl2 and mgcl2  (Millipore)


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    Name:
    Phosphate Buffered Saline
    Description:
    Phosphate Buffered Saline PBS is routinely used as a wash buffer in Western blot and immunoprecipitation IP procedures as well as during sample preparation of cell lysates for use in general molecular biology applications
    Catalog Number:
    p7059
    Price:
    None
    Applications:
    Suitable for use in wash buffers for Western blots and IP, as well as cell lysate sample preparation.
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    Structured Review

    Millipore phosphate buffered saline pbs without cacl2 and mgcl2
    Phosphate Buffered Saline PBS is routinely used as a wash buffer in Western blot and immunoprecipitation IP procedures as well as during sample preparation of cell lysates for use in general molecular biology applications
    https://www.bioz.com/result/phosphate buffered saline pbs without cacl2 and mgcl2/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phosphate buffered saline pbs without cacl2 and mgcl2 - by Bioz Stars, 2020-02
    95/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Role of Macromolecular Depletion in Red Blood Cell Adhesion
    Article Snippet: Blood was obtained by fingerstick using a sterile lancet and used within 4 h. RBCs were separated from whole blood by centrifugation at 1000 g for 5 min, and washed three times in isotonic phosphate-buffered saline solution (PBS, 10 mM including 0.2% bovine serum albumin). .. RBCs were then resuspended at 0.1% hematocrit in either PBS or dextran-PBS solutions: dextran 40 kDa, dextran 70 kDa, or dextran 500 kDa (Sigma Aldrich, Singapore) were dissolved in PBS at the desired final concentrations.

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs were reconstituted with 500 μL of 0.05 M carbonate/bicarbonate buffer (pH 9.5) and washed by centrifugation on vivaspin as described before.

    Immunocytochemistry:

    Article Title: Electrospun Nanofibers With pH-Responsive Coatings for Control of Release Kinetics
    Article Snippet: .. Immunocytochemistry of Coated PVA and Coated PVA-RB Nanofiber-Exposed Cells U87MG cells were washed with PBS with calcium and magnesium, and fixed with 4% paraformaldehyde (from Sigma-Aldrich) in PBS for 20 min at 4°C. .. Then, cells were again rinsed with PBS and permeabilized with 0.1% Triton X-100 (from Sigma-Aldrich) in PBS for 30 min.

    Positive Control:

    Article Title: Vaccination with the Recombinant Brucella Outer Membrane Protein 31 or a Derived 27-Amino-Acid Synthetic Peptide Elicits a CD4+ T Helper 1 Response That Protects against Brucella melitensis Infection
    Article Snippet: Mice were anesthetized with methoxyfuorane (Mallinckrodt, Phillipsburg, NJ) and immunized by the intraperitoneal (i.p.) route with 30 μg of rOmp31, 30 μg of Omp3148-74 , or phosphate-buffered saline (PBS) (as a control) in incomplete Freund's adjuvant (IFA) (Sigma, St. Louis, MO). .. As a positive control , another group was immunized once subcutaneously on day 0 with 8 × 108 formalin-killed B. melitensis H38S (H38) bacteria in IFA.

    Cytometry:

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Cell purity and phenotype of immature DCs were routinely determined by flow cytometry. .. Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate).

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: .. OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs were reconstituted with 500 μL of 0.05 M carbonate/bicarbonate buffer (pH 9.5) and washed by centrifugation on vivaspin as described before.

    Blocking Assay:

    Article Title: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma
    Article Snippet: Following 24 h of cell culture, the cells were treated with 4 µ M PAB for 24 h, washed with phosphate-buffered saline (PBS), fixed in 3.7% formaldehyde (Sigma-Aldrich) and rinsed three times in 1X PBS. .. The specimen was placed in blocking buffer (1X PBS, 5% normal serum and 0.3% Triton X-100) for 60 min, and subsequently incubated with α-tubulin antibody (1:100 dilution) overnight at 4°C.

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate). .. To block LPS effects, H. pylori sonicates were preincubated with 200 μg/ml of polymyxin B (Sigma-Aldrich) prior to the addition to DCs.

    CTL Assay:

    Article Title: Vaccination with the Recombinant Brucella Outer Membrane Protein 31 or a Derived 27-Amino-Acid Synthetic Peptide Elicits a CD4+ T Helper 1 Response That Protects against Brucella melitensis Infection
    Article Snippet: Mice were anesthetized with methoxyfuorane (Mallinckrodt, Phillipsburg, NJ) and immunized by the intraperitoneal (i.p.) route with 30 μg of rOmp31, 30 μg of Omp3148-74 , or phosphate-buffered saline (PBS) (as a control) in incomplete Freund's adjuvant (IFA) (Sigma, St. Louis, MO). .. Thirty days after the last protein injection, the mice were challenged intravenously with virulent Brucella organisms (eight mice per group) or were sacrificed to conduct the analysis of immune responses, including cytokine production and CTL induction (five mice per group).

    Incubation:

    Article Title: Electrospun Nanofibers With pH-Responsive Coatings for Control of Release Kinetics
    Article Snippet: Immunocytochemistry of Coated PVA and Coated PVA-RB Nanofiber-Exposed Cells U87MG cells were washed with PBS with calcium and magnesium, and fixed with 4% paraformaldehyde (from Sigma-Aldrich) in PBS for 20 min at 4°C. .. Then, cells were incubated with a 10% GS solution containing a rabbit polyclonal IgG primary antibody against Ki-67 (from Millipore, 1:100 diluted) for 3 h at 37°C.

    Article Title: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma
    Article Snippet: Following 24 h of cell culture, the cells were treated with 4 µ M PAB for 24 h, washed with phosphate-buffered saline (PBS), fixed in 3.7% formaldehyde (Sigma-Aldrich) and rinsed three times in 1X PBS. .. The specimen was placed in blocking buffer (1X PBS, 5% normal serum and 0.3% Triton X-100) for 60 min, and subsequently incubated with α-tubulin antibody (1:100 dilution) overnight at 4°C.

    Article Title: Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation
    Article Snippet: The mixtures contained 1% (w/v) pullulan, glycogen type IX, amylose, amylopectin or soluble starch (Sigma) dissolved in PBS (pH 7.0), and appropriately diluted enzyme in a total volume of 500 µL. .. After incubation at 37°C for 1 h, the reaction was stopped by addition of 1 mL of cold DNS buffer, followed by boiling for 15 min. 330 µL of a 40% potassium sodium tartrate (Rochelle salt) solution was added to each tubes to stabilize the color.

    Article Title: N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals
    Article Snippet: Then, 0.25% Triton® X-100 and 2 μl of PNGase F in deionized water were added and incubated at 37 °C for 24 h. Alternatively, 1 mg of each hemocyanin was dialyzed against dissociation buffer (130 mm glycine containing 2.5 mm EDTA, pH 8.6), and then heated at 60 or 100 °C for 15 min. Hemocyanins were cooled to room temperature, and 2 μl of PNGase F in deionized water were added and incubated for 24 h at 37 °C. .. After both deglycosylation methods, proteins were dialyzed against PBS or Tris buffer containing 5 mm MgCl2 and 5 mm CaCl2 , pH 7.2, in Amicon Ultra-15 10K MWCO tubes (Millipore).

    Article Title: Telomerase Enzyme Inhibition (TEI) and Cytolytic Therapy in the Management of Androgen Independent Osseous Metastatic Prostate Cancer
    Article Snippet: .. Tissue sections with either primary PSA antibody or PBS alone were inoculated at 4°C overnight in a humidified chamber, then washed with 0.05% Tween 20 (Sigma–Aldrich) in PBS 3 times and incubated with biotin-conjugated goat anti-rabbit IgG (Sigma–Aldrich) at room temperature for 60 min. After washing 3 times with 0.05% PBS-Tween 20, VECTOR-STAIN Elite ABC reagent (Vector Laboratories, Burlin-game, CA) was applied on tissues for 30 min at room temperature and brown color positive staining was developed with the Sigma FAST 3-3′-diaminobenzi-dine tablet set at room temperature for 10 min. .. The reaction was stopped by gently flushing with tap water for 10 min. Harris hematoxylin (Sigma–Aldrich) was used for counter-staining and cover glass was mounted with Permount (Biomeda, Foster City, CA).

    Activity Assay:

    Article Title: Functional Characterization of a Newly Identified Group B Streptococcus Pullulanase Eliciting Antibodies Able to Prevent Alpha-Glucans Degradation
    Article Snippet: 3,5-dinitrosalicylic acid (DNS) assay Pullulanase activity was determined by measuring the enzymatic release of reducing groups from α-glucans by the DNS colorimetric method . .. The mixtures contained 1% (w/v) pullulan, glycogen type IX, amylose, amylopectin or soluble starch (Sigma) dissolved in PBS (pH 7.0), and appropriately diluted enzyme in a total volume of 500 µL.

    Article Title: CP30, a Cysteine Proteinase Involved in Trichomonas vaginalis Cytoadherence
    Article Snippet: Before lysis, parasites were treated with different proteinase inhibitors to determine their effect on the CP30 proteolytic activity by substrate gel electrophoresis as before ( ). .. Briefly, 2 × 107 parasites suspended in phosphate-buffered saline (PBS) were treated for 20 min at 4°C with l -3-carboxy-2,3- trans -epoxypropionyl-leucylamido(4-guanidino)butane (E-64), p -tosyl- l -lysine chloromethyl ketone (TLCK), and leupeptin for cysteine proteinases; for serine proteinases, phenylmethylsulfonyl fluoride (PMSF); and for metalloproteinases, EDTA and EGTA (all purchased from Sigma Chemical Co., St. Louis, Mo.).

    Cell Culture:

    Article Title: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma
    Article Snippet: .. Following 24 h of cell culture, the cells were treated with 4 µ M PAB for 24 h, washed with phosphate-buffered saline (PBS), fixed in 3.7% formaldehyde (Sigma-Aldrich) and rinsed three times in 1X PBS. .. The specimen was placed in blocking buffer (1X PBS, 5% normal serum and 0.3% Triton X-100) for 60 min, and subsequently incubated with α-tubulin antibody (1:100 dilution) overnight at 4°C.

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Paragraph title: Cell culture. ... Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate).

    Article Title: The midgut transcriptome of Aedes aegypti fed with saline or protein meals containing chikungunya virus reveals genes potentially involved in viral midgut escape
    Article Snippet: .. Thereafter, cell culture media was collected and mixed with an equal amount of defibrinated sheep blood, 40% BSA (Sigma Aldrich, St. Louis, MO) or 2x PBS (pH 7.4, Sigma) including 10 mM ATP. .. One week-old females that had been deprived of sugar for 24 h were fed for 1 h with a mixture consisting of CHIKV-infected cell culture and blood, BSA, or PBS (or non-infected cell culture mixture as a control) at 37°C using a single glass feeder per carton.

    Expressing:

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Expression of CD14 was low and markedly downregulated compared to that of freshly isolated monocytes. .. Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate).

    Flow Cytometry:

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Cell purity and phenotype of immature DCs were routinely determined by flow cytometry. .. Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate).

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: .. OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs were reconstituted with 500 μL of 0.05 M carbonate/bicarbonate buffer (pH 9.5) and washed by centrifugation on vivaspin as described before.

    Protease Inhibitor:

    Article Title: Angiotensin-(1-7) inhibits allergic inflammation, via the MAS1 receptor, through suppression of ERK1/2- and NF-?B-dependent pathways
    Article Snippet: .. Chemicals used included ovalbumin (grade V), halothane, absolute ethanol (Merck KGaA, Darmstadt, Germany), formaldehyde (Surechem Products LTD, Suffolk, UK), Alu-Gel-S (SERVA Electrophoresis GmbH, Heidelberg, Germany), isotone II diluent solution (Beckman Coulter Inc., Krefeld, Germany), Zap-OGLOBIN (Coulter Electronics LTD, Buckinghamshire, UK), Diff-Quik (Baxter Dade AG, Dudingen, Switzerland), PBS (Sigma-Aldrich, St Louis, MO, USA), Ang-(1–7), A779, Tris-base, NaCl, Na4 P2 O7 , NaF, CaCl2 , MgCl2 , Glycerol, NP4 O, Na3 VO4 , PMSF, protease inhibitor cocktail, Ponceau 2R (Sigma-Aldrich), phosphatase inhibitor cocktail (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) xylene, ethanol (Honeywell Riedel-de Haën®, Seelze, Germany), haematoxylin, eosin, glacial acetic acid, ferric chloride, HCl, periodic acid, DMSO (BDH Laboratory Supplies, Poole, UK), Dpx mountant, picric acid, mercuric chloride, phosphomolybdic acid, light green SF yellowish, Schiff's reagent (Sigma-Aldrich), acid fuchsin (Gurr Microscopy Materials; BDH Ltd., Poole, UK), rabbit monoclonal antibodies for total and phosphorylated IκB-α and rabbit polyclonal antibody for phosphorylated ERK1/2 (Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-actin antibody, phenanthroline (Sigma-Aldrich), nitrocellulose membranes (Schleicher & Schuell, Dassel, Germany). .. Male BALB/c mice (6–8 weeks old) used in this study were maintained under temperature-controlled conditions with an artificial 12 h light/dark cycle and were allowed standard chow and water ad libitum .

    Infection:

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate). .. Bacteria were added either directly to the DCs at a multiplicity of infection (MOI) of 10 or to the top chambers of trans-wells (0.4-μm pore; Nunc GmbH Co., Wiesbaden, Germany) at an MOI of 100.

    Article Title: The midgut transcriptome of Aedes aegypti fed with saline or protein meals containing chikungunya virus reveals genes potentially involved in viral midgut escape
    Article Snippet: Paragraph title: CHIKV infection of mosquitoes and virus detection ... Thereafter, cell culture media was collected and mixed with an equal amount of defibrinated sheep blood, 40% BSA (Sigma Aldrich, St. Louis, MO) or 2x PBS (pH 7.4, Sigma) including 10 mM ATP.

    other:

    Article Title: Development of a β-Lactoglobulin Sensor Based on SPR for Milk Allergens Detection
    Article Snippet: Phosphate-buffered saline (PBS), β-lactoglobulin (BLG) from bovine milk, bovine serum albumin (BSA), sodium acetate, 11-mercaptoundecanoic acid 95% (MUDA), and sulphuric acid 95.0–98.0% were purchased from Sigma Aldrich (Saint Louis, MO, USA).

    Protein Concentration:

    Article Title: N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals
    Article Snippet: After both deglycosylation methods, proteins were dialyzed against PBS or Tris buffer containing 5 mm MgCl2 and 5 mm CaCl2 , pH 7.2, in Amicon Ultra-15 10K MWCO tubes (Millipore). .. The final protein concentration was determined by the Pierce® 660-nm colorimetric method (Thermo Fisher Scientific) according to the manufacturer's instructions.

    Injection:

    Article Title: Vaccination with the Recombinant Brucella Outer Membrane Protein 31 or a Derived 27-Amino-Acid Synthetic Peptide Elicits a CD4+ T Helper 1 Response That Protects against Brucella melitensis Infection
    Article Snippet: Mice were anesthetized with methoxyfuorane (Mallinckrodt, Phillipsburg, NJ) and immunized by the intraperitoneal (i.p.) route with 30 μg of rOmp31, 30 μg of Omp3148-74 , or phosphate-buffered saline (PBS) (as a control) in incomplete Freund's adjuvant (IFA) (Sigma, St. Louis, MO). .. Thirty days after the last protein injection, the mice were challenged intravenously with virulent Brucella organisms (eight mice per group) or were sacrificed to conduct the analysis of immune responses, including cytokine production and CTL induction (five mice per group).

    Binding Assay:

    Article Title: Electrospun Nanofibers With pH-Responsive Coatings for Control of Release Kinetics
    Article Snippet: Immunocytochemistry of Coated PVA and Coated PVA-RB Nanofiber-Exposed Cells U87MG cells were washed with PBS with calcium and magnesium, and fixed with 4% paraformaldehyde (from Sigma-Aldrich) in PBS for 20 min at 4°C. .. Specific binding sites were saturated with 10% goat serum (GS, from Thermo Scientific) in PBS for 1 h at 37°C.

    Immunofluorescence:

    Article Title: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma
    Article Snippet: Paragraph title: Immunofluorescence ... Following 24 h of cell culture, the cells were treated with 4 µ M PAB for 24 h, washed with phosphate-buffered saline (PBS), fixed in 3.7% formaldehyde (Sigma-Aldrich) and rinsed three times in 1X PBS.

    Article Title: Vaccination with the Recombinant Brucella Outer Membrane Protein 31 or a Derived 27-Amino-Acid Synthetic Peptide Elicits a CD4+ T Helper 1 Response That Protects against Brucella melitensis Infection
    Article Snippet: .. Mice were anesthetized with methoxyfuorane (Mallinckrodt, Phillipsburg, NJ) and immunized by the intraperitoneal (i.p.) route with 30 μg of rOmp31, 30 μg of Omp3148-74 , or phosphate-buffered saline (PBS) (as a control) in incomplete Freund's adjuvant (IFA) (Sigma, St. Louis, MO). .. As a positive control , another group was immunized once subcutaneously on day 0 with 8 × 108 formalin-killed B. melitensis H38S (H38) bacteria in IFA.

    Article Title: The midgut transcriptome of Aedes aegypti fed with saline or protein meals containing chikungunya virus reveals genes potentially involved in viral midgut escape
    Article Snippet: Thereafter, cell culture media was collected and mixed with an equal amount of defibrinated sheep blood, 40% BSA (Sigma Aldrich, St. Louis, MO) or 2x PBS (pH 7.4, Sigma) including 10 mM ATP. .. CHIKV antigen was detected by immunofluorescence assays and virus titers of individual midguts and carcasses were determined by plaque assays as described before [ ].

    Nucleic Acid Electrophoresis:

    Article Title: CP30, a Cysteine Proteinase Involved in Trichomonas vaginalis Cytoadherence
    Article Snippet: Before lysis, parasites were treated with different proteinase inhibitors to determine their effect on the CP30 proteolytic activity by substrate gel electrophoresis as before ( ). .. Briefly, 2 × 107 parasites suspended in phosphate-buffered saline (PBS) were treated for 20 min at 4°C with l -3-carboxy-2,3- trans -epoxypropionyl-leucylamido(4-guanidino)butane (E-64), p -tosyl- l -lysine chloromethyl ketone (TLCK), and leupeptin for cysteine proteinases; for serine proteinases, phenylmethylsulfonyl fluoride (PMSF); and for metalloproteinases, EDTA and EGTA (all purchased from Sigma Chemical Co., St. Louis, Mo.).

    Isolation:

    Article Title: Impact of Helicobacter pylori Virulence Factors and Compounds on Activation and Maturation of Human Dendritic Cells
    Article Snippet: Expression of CD14 was low and markedly downregulated compared to that of freshly isolated monocytes. .. Cells were either stimulated with 10 μl PBS, 100 ng/ml E. coli LPS 055:B5 (Sigma-Aldrich, Taufkirchen, Germany), or different H. pylori strains (formalin- or heat-inactivated H. pylori strains, as well as sonicate).

    Labeling:

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: .. OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs were reconstituted with 500 μL of 0.05 M carbonate/bicarbonate buffer (pH 9.5) and washed by centrifugation on vivaspin as described before.

    Mouse Assay:

    Article Title: Vaccination with the Recombinant Brucella Outer Membrane Protein 31 or a Derived 27-Amino-Acid Synthetic Peptide Elicits a CD4+ T Helper 1 Response That Protects against Brucella melitensis Infection
    Article Snippet: .. Mice were anesthetized with methoxyfuorane (Mallinckrodt, Phillipsburg, NJ) and immunized by the intraperitoneal (i.p.) route with 30 μg of rOmp31, 30 μg of Omp3148-74 , or phosphate-buffered saline (PBS) (as a control) in incomplete Freund's adjuvant (IFA) (Sigma, St. Louis, MO). .. As a positive control , another group was immunized once subcutaneously on day 0 with 8 × 108 formalin-killed B. melitensis H38S (H38) bacteria in IFA.

    Article Title: Temporal Changes in Glutaredoxin 1 and Protein S-Glutathionylation in Allergic Airway Inflammation
    Article Snippet: .. Briefly, mice were immunized intraperitoneally with 200 μL phosphate-buffered saline (PBS) containing 50 μg OVA (Grade V; Sigma-Aldrich, St. Louis, MO) plus 4.0 mg aluminum hydroxide adjuvant (Imject Alum; Thermo Scientific, Rockford, IL) on days 0 and 7. .. Mice (5 per group) were challenged with inhaled allergen (2.5% OVA in PBS) for 20 min or with PBS alone (control group) on days 21, 22, and 23.

    Diff-Quik:

    Article Title: Angiotensin-(1-7) inhibits allergic inflammation, via the MAS1 receptor, through suppression of ERK1/2- and NF-?B-dependent pathways
    Article Snippet: .. Chemicals used included ovalbumin (grade V), halothane, absolute ethanol (Merck KGaA, Darmstadt, Germany), formaldehyde (Surechem Products LTD, Suffolk, UK), Alu-Gel-S (SERVA Electrophoresis GmbH, Heidelberg, Germany), isotone II diluent solution (Beckman Coulter Inc., Krefeld, Germany), Zap-OGLOBIN (Coulter Electronics LTD, Buckinghamshire, UK), Diff-Quik (Baxter Dade AG, Dudingen, Switzerland), PBS (Sigma-Aldrich, St Louis, MO, USA), Ang-(1–7), A779, Tris-base, NaCl, Na4 P2 O7 , NaF, CaCl2 , MgCl2 , Glycerol, NP4 O, Na3 VO4 , PMSF, protease inhibitor cocktail, Ponceau 2R (Sigma-Aldrich), phosphatase inhibitor cocktail (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) xylene, ethanol (Honeywell Riedel-de Haën®, Seelze, Germany), haematoxylin, eosin, glacial acetic acid, ferric chloride, HCl, periodic acid, DMSO (BDH Laboratory Supplies, Poole, UK), Dpx mountant, picric acid, mercuric chloride, phosphomolybdic acid, light green SF yellowish, Schiff's reagent (Sigma-Aldrich), acid fuchsin (Gurr Microscopy Materials; BDH Ltd., Poole, UK), rabbit monoclonal antibodies for total and phosphorylated IκB-α and rabbit polyclonal antibody for phosphorylated ERK1/2 (Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-actin antibody, phenanthroline (Sigma-Aldrich), nitrocellulose membranes (Schleicher & Schuell, Dassel, Germany). .. Male BALB/c mice (6–8 weeks old) used in this study were maintained under temperature-controlled conditions with an artificial 12 h light/dark cycle and were allowed standard chow and water ad libitum .

    Immunostaining:

    Article Title: Telomerase Enzyme Inhibition (TEI) and Cytolytic Therapy in the Management of Androgen Independent Osseous Metastatic Prostate Cancer
    Article Snippet: For PSA immunostaining, tissue sections were dewaxed and rehydrated, endogenous peroxidase was quenched with 3% H2 O2 in methanol at room temperature for 30 min. After washing 3 times with PBS, sections were blocked with 0.05% BSA and 0.2% (v/v) normal goat serum (Sigma–Aldrich) in PBS at room temperature for 30 min. A polycolonal rabbit anti-human PSA antibody (BioGenex, San Ramon, CA) was used as the primary antibody at a 1:200 dilution in phosphate buffered saline (PBS). .. Tissue sections with either primary PSA antibody or PBS alone were inoculated at 4°C overnight in a humidified chamber, then washed with 0.05% Tween 20 (Sigma–Aldrich) in PBS 3 times and incubated with biotin-conjugated goat anti-rabbit IgG (Sigma–Aldrich) at room temperature for 60 min. After washing 3 times with 0.05% PBS-Tween 20, VECTOR-STAIN Elite ABC reagent (Vector Laboratories, Burlin-game, CA) was applied on tissues for 30 min at room temperature and brown color positive staining was developed with the Sigma FAST 3-3′-diaminobenzi-dine tablet set at room temperature for 10 min.

    Staining:

    Article Title: Pseudolaric acid B inhibits proliferation in SW579 human thyroid squamous cell carcinoma
    Article Snippet: Following 24 h of cell culture, the cells were treated with 4 µ M PAB for 24 h, washed with phosphate-buffered saline (PBS), fixed in 3.7% formaldehyde (Sigma-Aldrich) and rinsed three times in 1X PBS. .. The secondary antibodies were aspirated and the cover slips were rinsed once in 1X PBS prior to staining with 5 mg/l Hoechst 33258 for 30 min.

    Article Title: Telomerase Enzyme Inhibition (TEI) and Cytolytic Therapy in the Management of Androgen Independent Osseous Metastatic Prostate Cancer
    Article Snippet: .. Tissue sections with either primary PSA antibody or PBS alone were inoculated at 4°C overnight in a humidified chamber, then washed with 0.05% Tween 20 (Sigma–Aldrich) in PBS 3 times and incubated with biotin-conjugated goat anti-rabbit IgG (Sigma–Aldrich) at room temperature for 60 min. After washing 3 times with 0.05% PBS-Tween 20, VECTOR-STAIN Elite ABC reagent (Vector Laboratories, Burlin-game, CA) was applied on tissues for 30 min at room temperature and brown color positive staining was developed with the Sigma FAST 3-3′-diaminobenzi-dine tablet set at room temperature for 10 min. .. The reaction was stopped by gently flushing with tap water for 10 min. Harris hematoxylin (Sigma–Aldrich) was used for counter-staining and cover glass was mounted with Permount (Biomeda, Foster City, CA).

    Histone Association Assay:

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: .. OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs were reconstituted with 500 μL of 0.05 M carbonate/bicarbonate buffer (pH 9.5) and washed by centrifugation on vivaspin as described before.

    Plasmid Preparation:

    Article Title: Telomerase Enzyme Inhibition (TEI) and Cytolytic Therapy in the Management of Androgen Independent Osseous Metastatic Prostate Cancer
    Article Snippet: .. Tissue sections with either primary PSA antibody or PBS alone were inoculated at 4°C overnight in a humidified chamber, then washed with 0.05% Tween 20 (Sigma–Aldrich) in PBS 3 times and incubated with biotin-conjugated goat anti-rabbit IgG (Sigma–Aldrich) at room temperature for 60 min. After washing 3 times with 0.05% PBS-Tween 20, VECTOR-STAIN Elite ABC reagent (Vector Laboratories, Burlin-game, CA) was applied on tissues for 30 min at room temperature and brown color positive staining was developed with the Sigma FAST 3-3′-diaminobenzi-dine tablet set at room temperature for 10 min. .. The reaction was stopped by gently flushing with tap water for 10 min. Harris hematoxylin (Sigma–Aldrich) was used for counter-staining and cover glass was mounted with Permount (Biomeda, Foster City, CA).

    Solubility:

    Article Title: N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals
    Article Snippet: As the first protocol removed most N- glycans but decreased protein solubility, the second protocol was used for the subsequent experiments. .. After both deglycosylation methods, proteins were dialyzed against PBS or Tris buffer containing 5 mm MgCl2 and 5 mm CaCl2 , pH 7.2, in Amicon Ultra-15 10K MWCO tubes (Millipore).

    Microscopy:

    Article Title: Angiotensin-(1-7) inhibits allergic inflammation, via the MAS1 receptor, through suppression of ERK1/2- and NF-?B-dependent pathways
    Article Snippet: .. Chemicals used included ovalbumin (grade V), halothane, absolute ethanol (Merck KGaA, Darmstadt, Germany), formaldehyde (Surechem Products LTD, Suffolk, UK), Alu-Gel-S (SERVA Electrophoresis GmbH, Heidelberg, Germany), isotone II diluent solution (Beckman Coulter Inc., Krefeld, Germany), Zap-OGLOBIN (Coulter Electronics LTD, Buckinghamshire, UK), Diff-Quik (Baxter Dade AG, Dudingen, Switzerland), PBS (Sigma-Aldrich, St Louis, MO, USA), Ang-(1–7), A779, Tris-base, NaCl, Na4 P2 O7 , NaF, CaCl2 , MgCl2 , Glycerol, NP4 O, Na3 VO4 , PMSF, protease inhibitor cocktail, Ponceau 2R (Sigma-Aldrich), phosphatase inhibitor cocktail (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) xylene, ethanol (Honeywell Riedel-de Haën®, Seelze, Germany), haematoxylin, eosin, glacial acetic acid, ferric chloride, HCl, periodic acid, DMSO (BDH Laboratory Supplies, Poole, UK), Dpx mountant, picric acid, mercuric chloride, phosphomolybdic acid, light green SF yellowish, Schiff's reagent (Sigma-Aldrich), acid fuchsin (Gurr Microscopy Materials; BDH Ltd., Poole, UK), rabbit monoclonal antibodies for total and phosphorylated IκB-α and rabbit polyclonal antibody for phosphorylated ERK1/2 (Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit monoclonal anti-actin antibody, phenanthroline (Sigma-Aldrich), nitrocellulose membranes (Schleicher & Schuell, Dassel, Germany). .. Male BALB/c mice (6–8 weeks old) used in this study were maintained under temperature-controlled conditions with an artificial 12 h light/dark cycle and were allowed standard chow and water ad libitum .

    Negative Control:

    Article Title: Telomerase Enzyme Inhibition (TEI) and Cytolytic Therapy in the Management of Androgen Independent Osseous Metastatic Prostate Cancer
    Article Snippet: PBS without antibody was used as negative control. .. Tissue sections with either primary PSA antibody or PBS alone were inoculated at 4°C overnight in a humidified chamber, then washed with 0.05% Tween 20 (Sigma–Aldrich) in PBS 3 times and incubated with biotin-conjugated goat anti-rabbit IgG (Sigma–Aldrich) at room temperature for 60 min. After washing 3 times with 0.05% PBS-Tween 20, VECTOR-STAIN Elite ABC reagent (Vector Laboratories, Burlin-game, CA) was applied on tissues for 30 min at room temperature and brown color positive staining was developed with the Sigma FAST 3-3′-diaminobenzi-dine tablet set at room temperature for 10 min.

    Article Title: CP30, a Cysteine Proteinase Involved in Trichomonas vaginalis Cytoadherence
    Article Snippet: Briefly, 2 × 107 parasites suspended in phosphate-buffered saline (PBS) were treated for 20 min at 4°C with l -3-carboxy-2,3- trans -epoxypropionyl-leucylamido(4-guanidino)butane (E-64), p -tosyl- l -lysine chloromethyl ketone (TLCK), and leupeptin for cysteine proteinases; for serine proteinases, phenylmethylsulfonyl fluoride (PMSF); and for metalloproteinases, EDTA and EGTA (all purchased from Sigma Chemical Co., St. Louis, Mo.). .. As a negative control, parasites treated identically but in the absence of the proteinase inhibitors were used.

    Produced:

    Article Title: N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals
    Article Snippet: Chemically deglycosylated hemocyanins and OVA were produced by treatment with sodium periodate using the protocol described by Arancibia et al. ( ). .. After both deglycosylation methods, proteins were dialyzed against PBS or Tris buffer containing 5 mm MgCl2 and 5 mm CaCl2 , pH 7.2, in Amicon Ultra-15 10K MWCO tubes (Millipore).

    Concentration Assay:

    Article Title: Interspecies Outer Membrane Vesicles (OMVs) Modulate the Sensitivity of Pathogenic Bacteria and Pathogenic Yeasts to Cationic Peptides and Serum Complement
    Article Snippet: OMV Association Assay and Flow Cytometry OMVs labeling: Initially, OMVs (80 µg/mL) in PBS were concentrated using 50 kDa Vivaspin centrifugal concentrators (Amicon ultra, Millipore) at 14,000 × g for 10 min at 4 °C to remove PBS. .. The collected OMVs at concentration 80 µg/mL were labeled with 1 mg/mL FITC at carbonate/bicarbonate buffer for 30 min at 37 °C with gentle mixing in the dark.

    Lysis:

    Article Title: CP30, a Cysteine Proteinase Involved in Trichomonas vaginalis Cytoadherence
    Article Snippet: Before lysis, parasites were treated with different proteinase inhibitors to determine their effect on the CP30 proteolytic activity by substrate gel electrophoresis as before ( ). .. Briefly, 2 × 107 parasites suspended in phosphate-buffered saline (PBS) were treated for 20 min at 4°C with l -3-carboxy-2,3- trans -epoxypropionyl-leucylamido(4-guanidino)butane (E-64), p -tosyl- l -lysine chloromethyl ketone (TLCK), and leupeptin for cysteine proteinases; for serine proteinases, phenylmethylsulfonyl fluoride (PMSF); and for metalloproteinases, EDTA and EGTA (all purchased from Sigma Chemical Co., St. Louis, Mo.).

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    Millipore dulbecco s phosphate buffered saline
    Dulbecco S Phosphate Buffered Saline, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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