millipore pvdf membrane  (Millipore)


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    Name:
    Polyvinylidene difluoride membranes
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    Catalog Number:
    p0807
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    Structured Review

    Millipore millipore pvdf membrane
    Dependence of thermally on-off ratio on the cross-linker amounts in the range from 0.1-2.0 mol% in for <t>PNIPAAm-PVDF</t> <t>Millipore</t> membrane (P=1.4 bar). For all the membranes, the NIPAAm concentration for polymerization solution was 5 wt%. Data was corrected with viscosity and normalized by permeability at 30°C.

    https://www.bioz.com/result/millipore pvdf membrane/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    millipore pvdf membrane - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Development of Bench and Full-Scale Temperature and pH Responsive Functionalized PVDF Membranes with Tunable Properties"

    Article Title: Development of Bench and Full-Scale Temperature and pH Responsive Functionalized PVDF Membranes with Tunable Properties

    Journal: Journal of membrane science

    doi: 10.1016/j.memsci.2014.01.033

    Dependence of thermally on-off ratio on the cross-linker amounts in the range from 0.1-2.0 mol% in for PNIPAAm-PVDF Millipore membrane (P=1.4 bar). For all the membranes, the NIPAAm concentration for polymerization solution was 5 wt%. Data was corrected with viscosity and normalized by permeability at 30°C.
    Figure Legend Snippet: Dependence of thermally on-off ratio on the cross-linker amounts in the range from 0.1-2.0 mol% in for PNIPAAm-PVDF Millipore membrane (P=1.4 bar). For all the membranes, the NIPAAm concentration for polymerization solution was 5 wt%. Data was corrected with viscosity and normalized by permeability at 30°C.

    Techniques Used: Concentration Assay, Permeability

    ATR-FTIR spectrum of blank PVDF, PNIPAAm functionalized PVDF Millipore membrane and PNIPAAm-FPAA-PVDFHE Sepro membrane.
    Figure Legend Snippet: ATR-FTIR spectrum of blank PVDF, PNIPAAm functionalized PVDF Millipore membrane and PNIPAAm-FPAA-PVDFHE Sepro membrane.

    Techniques Used:

    Dynamic and Reversible flux response versus ramp change in feed temperature above LCST and below LCST through PNIPAAm-PVDF Millipore membrane at P = 1.4 bar. The inset is the experimental temperature step change approximation. For the polymerization, the NIPAAm concentration was 5 wt%, cross-linker was 0.1mol%.
    Figure Legend Snippet: Dynamic and Reversible flux response versus ramp change in feed temperature above LCST and below LCST through PNIPAAm-PVDF Millipore membrane at P = 1.4 bar. The inset is the experimental temperature step change approximation. For the polymerization, the NIPAAm concentration was 5 wt%, cross-linker was 0.1mol%.

    Techniques Used: Concentration Assay

    The effects of temperature and monomer concentration on dextran rejection with PNIPAAm-PVDF Millipore membrane (5mol% cross-linker) (Mw=2,000,000 g/mol; Stokes radius r s = 26.1nm, calculated from r s =0.27·M w 0.498 ).
    Figure Legend Snippet: The effects of temperature and monomer concentration on dextran rejection with PNIPAAm-PVDF Millipore membrane (5mol% cross-linker) (Mw=2,000,000 g/mol; Stokes radius r s = 26.1nm, calculated from r s =0.27·M w 0.498 ).

    Techniques Used: Concentration Assay

    SEM images of blank PVDF (A) and PNIPAAm-PVDF Millipore membranes (B: 25°C(below LCST); C: 40 °C (above LCST))
    Figure Legend Snippet: SEM images of blank PVDF (A) and PNIPAAm-PVDF Millipore membranes (B: 25°C(below LCST); C: 40 °C (above LCST))

    Techniques Used:

    Effect of monomer (NIPAAm) concentration on water flux at 1.4 bar and calculated effective pore size for PNIPAAm-PVDF Millipore membrane. cross-linker concentration = 1mol%.
    Figure Legend Snippet: Effect of monomer (NIPAAm) concentration on water flux at 1.4 bar and calculated effective pore size for PNIPAAm-PVDF Millipore membrane. cross-linker concentration = 1mol%.

    Techniques Used: Concentration Assay

    2) Product Images from "Pd/Fe nanoparticle integrated PMAA-PVDF membranes for chloro-organic remediation from synthetic and site groundwater"

    Article Title: Pd/Fe nanoparticle integrated PMAA-PVDF membranes for chloro-organic remediation from synthetic and site groundwater

    Journal: Journal of membrane science

    doi: 10.1016/j.memsci.2019.117454

    The SEM images for nanoparticles in the pores of membranes (PMAA functionalized Millipore DVPP04700 membranes). (a) An 80 μm membrane cross-section sample, the smooth area in the center, was prepared using FIB. (b) pore structure. (c) The nanoparticles inside the pores. (d) The quantification of particle size and distribution at various depths underneath the membrane surface (more than 300 particles were counted in every point). The Millipore PVDF membranes were used only for characterization.
    Figure Legend Snippet: The SEM images for nanoparticles in the pores of membranes (PMAA functionalized Millipore DVPP04700 membranes). (a) An 80 μm membrane cross-section sample, the smooth area in the center, was prepared using FIB. (b) pore structure. (c) The nanoparticles inside the pores. (d) The quantification of particle size and distribution at various depths underneath the membrane surface (more than 300 particles were counted in every point). The Millipore PVDF membranes were used only for characterization.

    Techniques Used:

    3) Product Images from "Pd/Fe nanoparticle integrated PMAA-PVDF membranes for chloro-organic remediation from synthetic and site groundwater"

    Article Title: Pd/Fe nanoparticle integrated PMAA-PVDF membranes for chloro-organic remediation from synthetic and site groundwater

    Journal: Journal of membrane science

    doi: 10.1016/j.memsci.2019.117454

    The SEM images for nanoparticles in the pores of membranes (PMAA functionalized Millipore DVPP04700 membranes). (a) An 80 μm membrane cross-section sample, the smooth area in the center, was prepared using FIB. (b) pore structure. (c) The nanoparticles inside the pores. (d) The quantification of particle size and distribution at various depths underneath the membrane surface (more than 300 particles were counted in every point). The Millipore PVDF membranes were used only for characterization.
    Figure Legend Snippet: The SEM images for nanoparticles in the pores of membranes (PMAA functionalized Millipore DVPP04700 membranes). (a) An 80 μm membrane cross-section sample, the smooth area in the center, was prepared using FIB. (b) pore structure. (c) The nanoparticles inside the pores. (d) The quantification of particle size and distribution at various depths underneath the membrane surface (more than 300 particles were counted in every point). The Millipore PVDF membranes were used only for characterization.

    Techniques Used:

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: 5-Keto-d-Gluconate Production Is Catalyzed by a Quinoprotein Glycerol Dehydrogenase, Major Polyol Dehydrogenase, in Gluconobacter Species
    Article Snippet: .. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed, proteins in the gel were transferred electrophoretically onto a polyvinylidene fluoride (PVDF) membrane (Millipore) at 100 mA for 4 h. After blocking with 3% gelatin and washing with 20 mM Tris-HCl buffer (pH 7.5) containing 500 mM NaCl (TBS), the membrane was incubated with anti-SLDH and also with anti-ARDH antibodies in TBS containing 1% gelatin for 2 h. Then, the membrane was washed again and incubated with TBS containing protein A-peroxidase for 2 h. After washing the membrane with TBS containing 0.05% Tween 20, the enzyme band was visualized by the addition of color reagents and H2 O2 . .. Prestained marker proteins (Bio-Rad) were used to estimate the relative molecular weight.

    Transfection:

    Article Title: Subcellular localization and expression of bamboo mosaic virus satellite RNA-encoded protein
    Article Snippet: .. Total proteins were extracted from transfected protoplasts or infected leaves , separated by 12.5 % SDS-PAGE and electrotransferred to PVDF membranes (Immobilon-P; Millipore). .. The blot was treated with rabbit polyclonal anti-P20 ( ) or anti-BaMV CP ( ) serum, or with mAb-P20, mAb-N20 or mAb-C21, followed by incubation with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG or anti-mouse IgG.

    Infection:

    Article Title: Subcellular localization and expression of bamboo mosaic virus satellite RNA-encoded protein
    Article Snippet: .. Total proteins were extracted from transfected protoplasts or infected leaves , separated by 12.5 % SDS-PAGE and electrotransferred to PVDF membranes (Immobilon-P; Millipore). .. The blot was treated with rabbit polyclonal anti-P20 ( ) or anti-BaMV CP ( ) serum, or with mAb-P20, mAb-N20 or mAb-C21, followed by incubation with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG or anti-mouse IgG.

    Electrophoresis:

    Article Title: Plastidial Starch Phosphorylase in Sweet Potato Roots Is Proteolytically Modified by Protein-Protein Interaction with the 20S Proteasome
    Article Snippet: .. After electrophoresis, proteins were either stained with Coomassie Brilliant Blue R-250 or transferred to the PVDF membrane (Immobilon-P, Millipore) for western blotting. .. In-gel Pho1 Activity Assay Samples were first separated on a 6% native PAGE.

    Article Title: Conditional Disruption of the Peroxisome Proliferator-Activated Receptor ? Gene in Mice Results in Lowered Expression of ABCA1, ABCG1, and apoE in Macrophages and Reduced Cholesterol Efflux
    Article Snippet: .. Then, 10 μg of total protein from Hepa-1 cells and 10 μg of protein from nuclear extracts of macrophages were subjected to electrophoresis on a 4 to 15% Tris-HCl gradient gel (Bio-Rad, Hercules, Calif.), transferred to Immobilon-P membranes (Millipore, Bedford, Mass.), and probed according to the manufacturer's recommendations with anti-PPARγ antibodies (E-8 and H-100; Santa Cruz Biotechnology, Inc., Santa Cruz, Calif.) as indicated. .. Detection of immunoreactive proteins was done by an enhanced chemiluminescence blot detection system (Amersham, Inc., Arlington Heights, Ill.).

    Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells
    Article Snippet: .. Aliquots of protein (10 μg) were heated to 100°C in Tris-Glycine SDS sample buffer (63 mM Tris HCl, pH 7.4, 10% glycerol, 2% SDS, 0.0025% bromphenol blue) (Novex) and 5% β-mercaptoethanol (Sigma) and separated by SDS-PAGE electrophoresis on precast 8% Tris-Glycine gels (Novex) and transferred to Immobilon P membranes (Millipore, Bedford, MA). .. Membranes were stained with Ponceau stain (Sigma) to ensure homogenous transfer of proteins and to allow for accurate marking of the transferred 10-κDa ladder (Gibco BRL) for estimation of protein molecular weight.

    Incubation:

    Article Title: 5-Keto-d-Gluconate Production Is Catalyzed by a Quinoprotein Glycerol Dehydrogenase, Major Polyol Dehydrogenase, in Gluconobacter Species
    Article Snippet: .. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed, proteins in the gel were transferred electrophoretically onto a polyvinylidene fluoride (PVDF) membrane (Millipore) at 100 mA for 4 h. After blocking with 3% gelatin and washing with 20 mM Tris-HCl buffer (pH 7.5) containing 500 mM NaCl (TBS), the membrane was incubated with anti-SLDH and also with anti-ARDH antibodies in TBS containing 1% gelatin for 2 h. Then, the membrane was washed again and incubated with TBS containing protein A-peroxidase for 2 h. After washing the membrane with TBS containing 0.05% Tween 20, the enzyme band was visualized by the addition of color reagents and H2 O2 . .. Prestained marker proteins (Bio-Rad) were used to estimate the relative molecular weight.

    Article Title: The inhibition of the apoptosis pathway by the Coxiella burnetii effector protein CaeA requires the EK repetition motif, but is independent of survivin
    Article Snippet: .. After 6h incubation at 37°C in 5%CO2 samples were separated by SDS-PAGE and transferred to a PVDF membrane (Millipore). .. The membranes were probed with antibodies directed against Bcl-2 (2870), Bcl-xL (2764), Mcl-1 (4572), cleaved caspase 7 (9491), cleaved caspase 9 (9501), XIAP (2045) and survivin (2808) all from Cell Signaling, cleaved PARP (611038, BD Biosciences) and actin (A2066, Sigma-Aldrich).

    Blocking Assay:

    Article Title: 5-Keto-d-Gluconate Production Is Catalyzed by a Quinoprotein Glycerol Dehydrogenase, Major Polyol Dehydrogenase, in Gluconobacter Species
    Article Snippet: .. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed, proteins in the gel were transferred electrophoretically onto a polyvinylidene fluoride (PVDF) membrane (Millipore) at 100 mA for 4 h. After blocking with 3% gelatin and washing with 20 mM Tris-HCl buffer (pH 7.5) containing 500 mM NaCl (TBS), the membrane was incubated with anti-SLDH and also with anti-ARDH antibodies in TBS containing 1% gelatin for 2 h. Then, the membrane was washed again and incubated with TBS containing protein A-peroxidase for 2 h. After washing the membrane with TBS containing 0.05% Tween 20, the enzyme band was visualized by the addition of color reagents and H2 O2 . .. Prestained marker proteins (Bio-Rad) were used to estimate the relative molecular weight.

    Article Title: The small splice variant of HPV16 E6, E6*, reduces tumor formation in cervical carcinoma xenografts HPV16 E6* reduces tumor formation
    Article Snippet: .. After protein transfer to Immobilon P membranes (Millipore Corporation) and blocking the membrane with 1% BSA, primary antibodies were applied in TBST containing 1% BSA. .. After incubation with primary antibodies overnight at 4°C, membranes were washed with TBST.

    Staining:

    Article Title: Plastidial Starch Phosphorylase in Sweet Potato Roots Is Proteolytically Modified by Protein-Protein Interaction with the 20S Proteasome
    Article Snippet: .. After electrophoresis, proteins were either stained with Coomassie Brilliant Blue R-250 or transferred to the PVDF membrane (Immobilon-P, Millipore) for western blotting. .. In-gel Pho1 Activity Assay Samples were first separated on a 6% native PAGE.

    Western Blot:

    Article Title: Plastidial Starch Phosphorylase in Sweet Potato Roots Is Proteolytically Modified by Protein-Protein Interaction with the 20S Proteasome
    Article Snippet: .. After electrophoresis, proteins were either stained with Coomassie Brilliant Blue R-250 or transferred to the PVDF membrane (Immobilon-P, Millipore) for western blotting. .. In-gel Pho1 Activity Assay Samples were first separated on a 6% native PAGE.

    SDS Page:

    Article Title: Subcellular localization and expression of bamboo mosaic virus satellite RNA-encoded protein
    Article Snippet: .. Total proteins were extracted from transfected protoplasts or infected leaves , separated by 12.5 % SDS-PAGE and electrotransferred to PVDF membranes (Immobilon-P; Millipore). .. The blot was treated with rabbit polyclonal anti-P20 ( ) or anti-BaMV CP ( ) serum, or with mAb-P20, mAb-N20 or mAb-C21, followed by incubation with horseradish peroxidase (HRP)-conjugated anti-rabbit IgG or anti-mouse IgG.

    Article Title: Responsiveness to Transforming Growth Factor-β (TGF-β)-Mediated Growth Inhibition Is a Function of Membrane-Bound TGF-β Type II Receptor in Human Breast Cancer Cells
    Article Snippet: .. Aliquots of protein (10 μg) were heated to 100°C in Tris-Glycine SDS sample buffer (63 mM Tris HCl, pH 7.4, 10% glycerol, 2% SDS, 0.0025% bromphenol blue) (Novex) and 5% β-mercaptoethanol (Sigma) and separated by SDS-PAGE electrophoresis on precast 8% Tris-Glycine gels (Novex) and transferred to Immobilon P membranes (Millipore, Bedford, MA). .. Membranes were stained with Ponceau stain (Sigma) to ensure homogenous transfer of proteins and to allow for accurate marking of the transferred 10-κDa ladder (Gibco BRL) for estimation of protein molecular weight.

    Article Title: 5-Keto-d-Gluconate Production Is Catalyzed by a Quinoprotein Glycerol Dehydrogenase, Major Polyol Dehydrogenase, in Gluconobacter Species
    Article Snippet: .. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed, proteins in the gel were transferred electrophoretically onto a polyvinylidene fluoride (PVDF) membrane (Millipore) at 100 mA for 4 h. After blocking with 3% gelatin and washing with 20 mM Tris-HCl buffer (pH 7.5) containing 500 mM NaCl (TBS), the membrane was incubated with anti-SLDH and also with anti-ARDH antibodies in TBS containing 1% gelatin for 2 h. Then, the membrane was washed again and incubated with TBS containing protein A-peroxidase for 2 h. After washing the membrane with TBS containing 0.05% Tween 20, the enzyme band was visualized by the addition of color reagents and H2 O2 . .. Prestained marker proteins (Bio-Rad) were used to estimate the relative molecular weight.

    Article Title: The inhibition of the apoptosis pathway by the Coxiella burnetii effector protein CaeA requires the EK repetition motif, but is independent of survivin
    Article Snippet: .. After 6h incubation at 37°C in 5%CO2 samples were separated by SDS-PAGE and transferred to a PVDF membrane (Millipore). .. The membranes were probed with antibodies directed against Bcl-2 (2870), Bcl-xL (2764), Mcl-1 (4572), cleaved caspase 7 (9491), cleaved caspase 9 (9501), XIAP (2045) and survivin (2808) all from Cell Signaling, cleaved PARP (611038, BD Biosciences) and actin (A2066, Sigma-Aldrich).

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    Millipore immobilon p polyvinylidenedifluoride pvdf membranes
    Western blotting analysis of Halobacterium sp. NRC-1 andderivative strains using rabbit Rfa3 antiserum. A. Equal quantities of totalcell lysate protein samples were electrophoresed on a 12% polyacrylamide-SDSgel, transferred to <t>PVDF</t> membrane, and probed
    Immobilon P Polyvinylidenedifluoride Pvdf Membranes, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 759 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/immobilon p polyvinylidenedifluoride pvdf membranes/product/Millipore
    Average 99 stars, based on 759 article reviews
    Price from $9.99 to $1999.99
    immobilon p polyvinylidenedifluoride pvdf membranes - by Bioz Stars, 2020-09
    99/100 stars
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    Western blotting analysis of Halobacterium sp. NRC-1 andderivative strains using rabbit Rfa3 antiserum. A. Equal quantities of totalcell lysate protein samples were electrophoresed on a 12% polyacrylamide-SDSgel, transferred to PVDF membrane, and probed

    Journal: Applied microbiology and biotechnology

    Article Title: Bioengineering radioresistance by overproduction of RPA, a mammalian-type single-stranded DNA-binding protein, in a halophilic archaeon

    doi: 10.1007/s00253-013-5368-x

    Figure Lengend Snippet: Western blotting analysis of Halobacterium sp. NRC-1 andderivative strains using rabbit Rfa3 antiserum. A. Equal quantities of totalcell lysate protein samples were electrophoresed on a 12% polyacrylamide-SDSgel, transferred to PVDF membrane, and probed

    Article Snippet: Briefly, the electrophoretically fractionated cell lysates orpure protein were electroblotted onto 0.45 μm Immobilon-P polyvinylidenedifluoride (PVDF) membranes (Millipore Corp., Boston, MA) for 1 h at 100 voltsusing a Bio-Rad mini gel blotter.

    Techniques: Western Blot

    Distribution of tubulin in MDCK clone II/G and II/J cells after treatment with nocodazole. (A) Timeline scheme of experimental protocol used to disrupt microtubules. Confluent MDCK clone II/G and II/J cultures were established as described in MATERIALS AND METHODS. (B) After incubation in either the absence (−noc) or presence (+noc) of nocodazole, 120-h differentiated MDCK clone II/G and clone II/J cells were fixed and processed for immunofluorescence with monoclonal antibodies against α- and β-tubulin (see MATERIALS AND METHODS) to show the distribution of intact microtubules. Image stacks were produced from serial images collected for each specimen using a z step of 0.2 μm. (C) At the indicated times after the induction of cell–cell contacts, the amount of soluble (S) and polymerized tubulin (P) was determined in both clones of MDCK cells incubated in either the absence (−noc) or presence (+noc) of nocodazole. Samples were extracted in microtubule-stabilizing buffer containing 0.1% Triton X-100 and centrifuged at 20,000 × g to separate soluble (S) and insoluble fractions (P). The insoluble fractions were then solubilized in 0.5% Triton X-100. Equal amounts of supernatant and pellet fractions for each sample were separated by SDS-PAGE. Gels were then electrophoretically transferred to Immobilon polyvinylidene fluoride membrane and probed with a monoclonal antibody to β-tubulin. The β-tubulin antibody was visualized with a horseradish peroxidase-conjugated secondary antibody followed by enhanced chemiluminescence. Representative samples from two to four independent experiments for each time point are presented. Bar, 10 μm.

    Journal: Molecular Biology of the Cell

    Article Title: Apiconuclear Organization of Microtubules Does Not Specify Protein Delivery from the Trans-Golgi Network to Different Membrane Domains in Polarized Epithelial Cells

    doi:

    Figure Lengend Snippet: Distribution of tubulin in MDCK clone II/G and II/J cells after treatment with nocodazole. (A) Timeline scheme of experimental protocol used to disrupt microtubules. Confluent MDCK clone II/G and II/J cultures were established as described in MATERIALS AND METHODS. (B) After incubation in either the absence (−noc) or presence (+noc) of nocodazole, 120-h differentiated MDCK clone II/G and clone II/J cells were fixed and processed for immunofluorescence with monoclonal antibodies against α- and β-tubulin (see MATERIALS AND METHODS) to show the distribution of intact microtubules. Image stacks were produced from serial images collected for each specimen using a z step of 0.2 μm. (C) At the indicated times after the induction of cell–cell contacts, the amount of soluble (S) and polymerized tubulin (P) was determined in both clones of MDCK cells incubated in either the absence (−noc) or presence (+noc) of nocodazole. Samples were extracted in microtubule-stabilizing buffer containing 0.1% Triton X-100 and centrifuged at 20,000 × g to separate soluble (S) and insoluble fractions (P). The insoluble fractions were then solubilized in 0.5% Triton X-100. Equal amounts of supernatant and pellet fractions for each sample were separated by SDS-PAGE. Gels were then electrophoretically transferred to Immobilon polyvinylidene fluoride membrane and probed with a monoclonal antibody to β-tubulin. The β-tubulin antibody was visualized with a horseradish peroxidase-conjugated secondary antibody followed by enhanced chemiluminescence. Representative samples from two to four independent experiments for each time point are presented. Bar, 10 μm.

    Article Snippet: For tubulin samples, the gels were electrophoretically transferred to Immobilon polyvinylidene fluoride membrane (Millipore, Bedford, MA).

    Techniques: Incubation, Immunofluorescence, Produced, Clone Assay, SDS Page

    Two-dimensional gel electrophoresis patterns of the proteins-secreted into medium from  S. enterica  serovar Typhimurium CS2033 ( fljB ::Tn 10 ) (A), CS2034 ( fljB ::Tn 10 clpP ::Cm r ) (B), CS2144 ( fljB ::Tn 10 flhD-lac ) (C), and CS2145 ( fljB ::Tn 10 flhD-lac clpP ::Cm r ) (D). Protein spots excised for mass spectrometry analysis or transferred to the polyvinylidene difluoride membrane for amino-terminal sequence analysis are interpreted in the text.

    Journal: Journal of Bacteriology

    Article Title: The ClpXP ATP-Dependent Protease Regulates Flagellum Synthesis in Salmonella enterica Serovar Typhimurium

    doi: 10.1128/JB.184.3.645-653.2002

    Figure Lengend Snippet: Two-dimensional gel electrophoresis patterns of the proteins-secreted into medium from S. enterica serovar Typhimurium CS2033 ( fljB ::Tn 10 ) (A), CS2034 ( fljB ::Tn 10 clpP ::Cm r ) (B), CS2144 ( fljB ::Tn 10 flhD-lac ) (C), and CS2145 ( fljB ::Tn 10 flhD-lac clpP ::Cm r ) (D). Protein spots excised for mass spectrometry analysis or transferred to the polyvinylidene difluoride membrane for amino-terminal sequence analysis are interpreted in the text.

    Article Snippet: For immunoblot analysis, proteins separated on the SDS-12% polyacrylamide gels were transferred after electrophoresis onto Immobilon-P (Millipore) as previously reported ( ).

    Techniques: Two-Dimensional Gel Electrophoresis, Electrophoresis, Mass Spectrometry, Sequencing

    ( A ) Chemical properties of the phosphoenzyme intermediate.  32 P-labeled, phosphorylated Nuc was transferred to Immobilon P. Filter strips were counted, then incubated in: ( i ) 1 M HCl; ( ii ) 1 M NaOH; ( iii ) 1 M hydroxylamine in 200 mM Tris⋅Cl, pH 7.4; and ( iv )1 M Tris⋅Cl, pH 7.4, for 45 min at 37°C. Radioactivity remaining on each filter strip was visualized by autoradiography and quantitated by counting. ( B ) Filter strips were incubated in 50 mM sodium citrate, pH 2.5, or in solutions of 25 mM Tris, 25 mM Bis·Tris, and 50 mM acetic acid at pH values ranging from 3.5 to 8.8, for 30 min at 80°C. The amount of radioactivity on each filter strip before and after incubation was quantitated by scintillation spectrometry.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Catalytic mechanism of the phospholipase D superfamily proceeds via a covalent phosphohistidine intermediate

    doi:

    Figure Lengend Snippet: ( A ) Chemical properties of the phosphoenzyme intermediate. 32 P-labeled, phosphorylated Nuc was transferred to Immobilon P. Filter strips were counted, then incubated in: ( i ) 1 M HCl; ( ii ) 1 M NaOH; ( iii ) 1 M hydroxylamine in 200 mM Tris⋅Cl, pH 7.4; and ( iv )1 M Tris⋅Cl, pH 7.4, for 45 min at 37°C. Radioactivity remaining on each filter strip was visualized by autoradiography and quantitated by counting. ( B ) Filter strips were incubated in 50 mM sodium citrate, pH 2.5, or in solutions of 25 mM Tris, 25 mM Bis·Tris, and 50 mM acetic acid at pH values ranging from 3.5 to 8.8, for 30 min at 80°C. The amount of radioactivity on each filter strip before and after incubation was quantitated by scintillation spectrometry.

    Article Snippet: The gel was soaked in buffer containing 12.5 mM Tris base, 96 mM glycine, and 20% methanol, and the protein was transferred to an Immobilon P (Millipore) membrane by electrophoresis at 100 V for 1 hr in the same buffer.

    Techniques: Labeling, Incubation, Radioactivity, Stripping Membranes, Autoradiography