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Bio-Rad tris hcl polyacrylamide gels
Tris Hcl Polyacrylamide Gels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Polyacrylamide Gel Electrophoresis:

Article Title: Ixeris dentata extract regulates salivary secretion through the activation of aquaporin-5 and prevents diabetes-induced xerostomia
Article Snippet: The composition of lysis buffer was prepared as described in Lee et al. Total protein was quantified using the BioRad Protein assay (BioRad). .. Thirty µg of total protein were loaded per lane, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; BioRad), and transferred to a polyvinylidene fluoride membrane. ..

Article Title: Calcium Homeostasis and Muscle Energy Metabolism Are Modified in HspB1-Null Mice
Article Snippet: .. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (T = 11%, C = 2.6%) was run on a PROTEAN plus Dodeca cell system (Bio-Rad, Hercules, CA, USA) at 40 V for 1 h, with 15 mA per gel and 110 V until the Bromophenol blue migration front reached the bottom of all the gels. .. Three replicates of two-DE (Dimensional electrophoresis) gels of HspB1-null mice (n = 5 samples) and their control littermates (n = 5 samples) were carried out.

Article Title: Intracellular delivery of Parkin rescues neurons from accumulation of damaged mitochondria and pathological α-synuclein
Article Snippet: Western blot analysis and ELISA for α-synuclein clearance in animal studies To analyze α-synuclein clearance in brain samples, the isolated brains were homogenized in PRO-PREP lysis buffer (iNtRON Biotechnology) containing a protease inhibitor (Thermo Fisher Scientific). .. The quantified cell lysates were separated by 10% SDS–polyacrylamide gel electrophoresis (PAGE) and transferred to nitrocellulose membranes (Bio-Rad). .. Membranes were incubated with primary antibodies against α-synuclein (1:1000; Santa Cruz Biotechnology, SC-12767), TH (1:2000; Millipore, AB152), pSer129-α-synuclein (1:5000; Abcam, ab51253), and β-actin (1:100,000; Sigma-Aldrich, A3854) followed by secondary antibodies.

Article Title: LC-MS/MS Analysis of Canine Lipoproteins Fractionated Using the Ultracentrifugation-Precipitation Method
Article Snippet: .. Briefly, 7 microliters of the HDL+LDL, HDL and LDL fractions, separated using the U-P method, were subjected to SDS polyacrylamide gel electrophoresis (SDS-PAGE) with 4–20% gradient acrylamide gels (Ready Gel J, Bio-Rad Laboratories, Inc., Benicia, CA, U.S.A.) and stained with Coomassie Brilliant Blue [ ]. .. The stained protein’s bands were excised with a scalpel, washed with 100 µl of 50% acetonitrile in 25 mM ammonium carbonate and left to dry at RT for 10 min with shaking in an Eppendorf Thermomixer.

SDS Page:

Article Title: Ixeris dentata extract regulates salivary secretion through the activation of aquaporin-5 and prevents diabetes-induced xerostomia
Article Snippet: The composition of lysis buffer was prepared as described in Lee et al. Total protein was quantified using the BioRad Protein assay (BioRad). .. Thirty µg of total protein were loaded per lane, separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; BioRad), and transferred to a polyvinylidene fluoride membrane. ..

Article Title: Potent inhibitors of toxic alpha-synuclein oligomers identified via cellular time-resolved FRET biosensor
Article Snippet: Cells were lysed for 30 minutes on ice with radioimmunoprecipitation assay (RIPA) lysis buffer (Pierce RIPA buffer, Thermo Fisher Scientific) containing 1% protease inhibitor (Clontech, Mountain View, CA) and 1% phosphatase inhibitors (Millipore Sigma), and centrifuged at 15,000 g at 4 °C for 15 min. .. The total protein concentration of lysates was determined by bicinchoninic acid (BCA) assay (Pierce), and equal amounts of total protein (60 μg) were mixed with 4× Bio-Rad sample buffer and loaded onto 4%–15% Trisglycine sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels (Bio-Rad, Hercules, CA). .. Proteins were transferred to supported nitrocellulose membrane and probed using Syn101 antibodies against aSyn (BD labs, San Jose, CA).

Article Title: Calcium Homeostasis and Muscle Energy Metabolism Are Modified in HspB1-Null Mice
Article Snippet: .. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (T = 11%, C = 2.6%) was run on a PROTEAN plus Dodeca cell system (Bio-Rad, Hercules, CA, USA) at 40 V for 1 h, with 15 mA per gel and 110 V until the Bromophenol blue migration front reached the bottom of all the gels. .. Three replicates of two-DE (Dimensional electrophoresis) gels of HspB1-null mice (n = 5 samples) and their control littermates (n = 5 samples) were carried out.

Article Title: Biochemical and Biological Characterization of the Protective Leishmania pifanoi Amastigote Antigen P-8
Article Snippet: The fractions eluted from the affinity column were assessed for protein by measuring the absorbance at 280 and 320 nm; protein fractions were pooled, concentrated, and then stored at −20°C. .. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was carried out with a minigel system (Bio-Rad) and 12% acrylamide gels ( ). ..

Article Title: LC-MS/MS Analysis of Canine Lipoproteins Fractionated Using the Ultracentrifugation-Precipitation Method
Article Snippet: .. Briefly, 7 microliters of the HDL+LDL, HDL and LDL fractions, separated using the U-P method, were subjected to SDS polyacrylamide gel electrophoresis (SDS-PAGE) with 4–20% gradient acrylamide gels (Ready Gel J, Bio-Rad Laboratories, Inc., Benicia, CA, U.S.A.) and stained with Coomassie Brilliant Blue [ ]. .. The stained protein’s bands were excised with a scalpel, washed with 100 µl of 50% acetonitrile in 25 mM ammonium carbonate and left to dry at RT for 10 min with shaking in an Eppendorf Thermomixer.

Nucleic Acid Electrophoresis:

Article Title: 2,2,2-Trifluoroethanol Changes the Transition Kinetics and Subunit Interactions in the Small Bacterial Mechanosensitive Channel MscS
Article Snippet: Coomassie Brilliant Blue G-250 was purchased from ICN Biomedicals (Aurora, OH). .. Lithium dodecyl sulfate-polyacrylamide gel electrophoresis (LDS-PAGE) gradient gels were cast using a Hoefner SG30 gel maker while nongradient LDS gels were cast on BioRad Protean III casting systems. .. PB111, a plasmid containing MscS with a C-terminal 6 His tag, was a gift of Dr. Paul Blount (UT Southwestern, Dallas, TX).

Article Title: Potent inhibitors of toxic alpha-synuclein oligomers identified via cellular time-resolved FRET biosensor
Article Snippet: Cells were lysed for 30 minutes on ice with radioimmunoprecipitation assay (RIPA) lysis buffer (Pierce RIPA buffer, Thermo Fisher Scientific) containing 1% protease inhibitor (Clontech, Mountain View, CA) and 1% phosphatase inhibitors (Millipore Sigma), and centrifuged at 15,000 g at 4 °C for 15 min. .. The total protein concentration of lysates was determined by bicinchoninic acid (BCA) assay (Pierce), and equal amounts of total protein (60 μg) were mixed with 4× Bio-Rad sample buffer and loaded onto 4%–15% Trisglycine sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels (Bio-Rad, Hercules, CA). .. Proteins were transferred to supported nitrocellulose membrane and probed using Syn101 antibodies against aSyn (BD labs, San Jose, CA).

Article Title: Intracellular delivery of Parkin rescues neurons from accumulation of damaged mitochondria and pathological α-synuclein
Article Snippet: Western blot analysis and ELISA for α-synuclein clearance in animal studies To analyze α-synuclein clearance in brain samples, the isolated brains were homogenized in PRO-PREP lysis buffer (iNtRON Biotechnology) containing a protease inhibitor (Thermo Fisher Scientific). .. The quantified cell lysates were separated by 10% SDS–polyacrylamide gel electrophoresis (PAGE) and transferred to nitrocellulose membranes (Bio-Rad). .. Membranes were incubated with primary antibodies against α-synuclein (1:1000; Santa Cruz Biotechnology, SC-12767), TH (1:2000; Millipore, AB152), pSer129-α-synuclein (1:5000; Abcam, ab51253), and β-actin (1:100,000; Sigma-Aldrich, A3854) followed by secondary antibodies.

Protein Concentration:

Article Title: Potent inhibitors of toxic alpha-synuclein oligomers identified via cellular time-resolved FRET biosensor
Article Snippet: Cells were lysed for 30 minutes on ice with radioimmunoprecipitation assay (RIPA) lysis buffer (Pierce RIPA buffer, Thermo Fisher Scientific) containing 1% protease inhibitor (Clontech, Mountain View, CA) and 1% phosphatase inhibitors (Millipore Sigma), and centrifuged at 15,000 g at 4 °C for 15 min. .. The total protein concentration of lysates was determined by bicinchoninic acid (BCA) assay (Pierce), and equal amounts of total protein (60 μg) were mixed with 4× Bio-Rad sample buffer and loaded onto 4%–15% Trisglycine sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels (Bio-Rad, Hercules, CA). .. Proteins were transferred to supported nitrocellulose membrane and probed using Syn101 antibodies against aSyn (BD labs, San Jose, CA).

BIA-KA:

Article Title: Potent inhibitors of toxic alpha-synuclein oligomers identified via cellular time-resolved FRET biosensor
Article Snippet: Cells were lysed for 30 minutes on ice with radioimmunoprecipitation assay (RIPA) lysis buffer (Pierce RIPA buffer, Thermo Fisher Scientific) containing 1% protease inhibitor (Clontech, Mountain View, CA) and 1% phosphatase inhibitors (Millipore Sigma), and centrifuged at 15,000 g at 4 °C for 15 min. .. The total protein concentration of lysates was determined by bicinchoninic acid (BCA) assay (Pierce), and equal amounts of total protein (60 μg) were mixed with 4× Bio-Rad sample buffer and loaded onto 4%–15% Trisglycine sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) gels (Bio-Rad, Hercules, CA). .. Proteins were transferred to supported nitrocellulose membrane and probed using Syn101 antibodies against aSyn (BD labs, San Jose, CA).

Migration:

Article Title: Calcium Homeostasis and Muscle Energy Metabolism Are Modified in HspB1-Null Mice
Article Snippet: .. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (T = 11%, C = 2.6%) was run on a PROTEAN plus Dodeca cell system (Bio-Rad, Hercules, CA, USA) at 40 V for 1 h, with 15 mA per gel and 110 V until the Bromophenol blue migration front reached the bottom of all the gels. .. Three replicates of two-DE (Dimensional electrophoresis) gels of HspB1-null mice (n = 5 samples) and their control littermates (n = 5 samples) were carried out.

Purification:

Article Title: Modular DNA strand-displacement controllers for directing material expansion
Article Snippet: .. Acrylamide (Bio-Rad, Cat. No. 161-0100) was solubilized using MilliQ purified water. .. Rhodamine B-conjugated methacrylate monomer was obtained from PolySciences, Inc (Cat. No. 25404-100) and used for fluorescent visualization of hydrogels.

Staining:

Article Title: LC-MS/MS Analysis of Canine Lipoproteins Fractionated Using the Ultracentrifugation-Precipitation Method
Article Snippet: .. Briefly, 7 microliters of the HDL+LDL, HDL and LDL fractions, separated using the U-P method, were subjected to SDS polyacrylamide gel electrophoresis (SDS-PAGE) with 4–20% gradient acrylamide gels (Ready Gel J, Bio-Rad Laboratories, Inc., Benicia, CA, U.S.A.) and stained with Coomassie Brilliant Blue [ ]. .. The stained protein’s bands were excised with a scalpel, washed with 100 µl of 50% acetonitrile in 25 mM ammonium carbonate and left to dry at RT for 10 min with shaking in an Eppendorf Thermomixer.

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    Bio-Rad acrylamide tris hcl ready sds page gels
    Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by <t>SDS-PAGE</t> analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM <t>Hepes-Tris,</t> pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.
    Acrylamide Tris Hcl Ready Sds Page Gels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/acrylamide tris hcl ready sds page gels/product/Bio-Rad
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    96
    Bio-Rad criterion tris hcl precast sds page gels
    Subcellular localization of DmLRPPRC2. (A) Fluorescence confocal microscopy images showing HeLa cells ( n = 15) expressing a DmLRPPRC2-GFP fusion protein and counterstained with the mitochondrial marker MitoTracker Deep Red. Scale bar: 10 μm. (B) Subcellular fractionation showing the presence of DmLRPPRC2 in the mitochondrial fraction. Subcellular fractions were separated by <t>SDS-PAGE</t> followed by western blot analysis to detect histone H3 (a nuclear marker), tubulin (a cytosolic marker), VDAC (a mitochondrial marker) and DmLRPPRC2. See also Supplementary Figure S1A and B.
    Criterion Tris Hcl Precast Sds Page Gels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/criterion tris hcl precast sds page gels/product/Bio-Rad
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    criterion tris hcl precast sds page gels - by Bioz Stars, 2021-03
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    Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 1. Effect of LAs on temperature dependence of KcsA tetramer stability in the presence and absence of K + . KcsA tetramer was measured by SDS-PAGE analysis of a series of identical samples incubated at various temperatures for 10 min before addition of SDS-PAGE sample buffer. The sample assay mixture contained 10 mM Hepes-Tris, pH 7.4, 100 mM cholineCl, either 5 mM KCl ( A ) or no added KCl ( B ) and either no LA (○), 20 mM lidocaine (●) or 5 mM tetracaine (△). Solid lines indicate nonlinear regression fits to a logistic function of temperature described in Materials and Methods . Fit parameters: ( A ) 20 mM lidocaine (●) : T 0.5 = 98.0 ± 2.1 °C, n = 14.1 ± 4.8; ( A ) 5 mM tetracaine (△): T 0.5 = 75.9 ± 1.7 °C, n = 10.7 ± 2.3; ( B ) no LA (○): T 0.5 = 45.7 ± 0.9 °C, n = 6.4 ± 0.7; ( B ) 20 mM lidocaine (●) : T 0.5 = 41.9 ± 0.7 °C, n = 6.4 ± 0.6.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: SDS Page, Incubation

    Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 2. Destabilization of KcsA tetramer as function of LA concentration. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing concentrations of lidocaine (●) or tetracaine (△, □ ). The sample assay mixture contained 10 mM Hepes-Tris, pH 7.5, 100 mM cholineCl and either 5 mM KCl (● , △) or no added KCl (□ ). Samples were incubated either at 90°C (● , △) or 22 °C (□ ) for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of LA concentration as described in Materials and Methods . Fit parameters: lidocaine, 5 mM KCl, 90°C (● ): IC 50 = 25.1 ± 2.6 mM, n = 1.14 ± 0.13; tetracaine, 5 mM KCl, 90 °C (△): IC 50 = 4.2 ± 0.6 mM, n = 1.43 ± 0.33; tetracaine, 0 KCl, 22 °C (□ ): 1.2 ± 0.2 mM, n = 0.98 ± 0.22.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: Concentration Assay, SDS Page, Incubation

    Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.

    Journal: Channels

    Article Title: Interaction of local anesthetics with the K+ channel pore domain

    doi: 10.4161/chan.24455

    Figure Lengend Snippet: Figure 3. Stabilization of KcsA tetramer as a function of K + concentration in the absence and presence of LA. KcsA tetramer was measured by SDS-PAGE analysis of samples titrated with increasing K + concentration in the absence of LA (○) or in the presence of 20 mM lidocaine (● ) or 5 mM tetracaine (△). The sample assay mixture contained ~500 ng KcsA, 10 mM Hepes-Tris, pH 7.4, 100 mM choline Cl, indicated concentrations of KCl and either no LA, 20 mM lidocaine or 5 mM tetracaine. All samples were incubated at 90 °C for 10 min before addition of SDS-PAGE sample buffer. Solid lines indicate nonlinear regression fits to a logistic function of K + concentration as described in Materials and Methods . Fit parameters: no LA (○): K 0.5 = 1.48 ± 0.04 mM, n = 3.19 ± 0.26; 20 mM lidocaine (● ): K 0.5 = 6.65 ± 0.8 mM, n = 1.05 ± 0.13; 5 mM tetracaine (△): K 0.5 = 5.03 ± 0.90, n = 1.04 ± 0.19.

    Article Snippet: Ten milliliters of each sample was pipetted into wells of precast 12% acrylamide TRIS-HCl ready SDS-PAGE Gels (Bio-Rad) and subjected to electrophoresis at 200V for ~35 min in running buffer (25 mM Tris base, 192 mM glycine, pH 8.3, 0.1% SDS).

    Techniques: Concentration Assay, SDS Page, Incubation

    Cystatin A inhibits the processing of MYOC wild-type in cultured cells. Recombinant expression plasmids containing tag-fused full coding wild-type MYOC (pMG29), CSTA, and controls plasmids, inactive mutated CSTA (CSTAm) and pEmpty, were generated as indicated in Methods . pMG29 was co-transfected with either pCSTA, pCSTAm or pEmpty (1∶2) and harvested at 48 h post-transfection. Equivalent volumes of cell extracts and of their supernatants were loaded onto 4–15% SDS-PAGE gels, transferred to PVDF membranes and analyzed by immunoblotting. Different MYOC protein forms (full length and processed) were detected with an anti-V5 mouse monoclonal followed by an anti-mouse horseradish peroxidase antibodies. Blots were re-probed with β-actin and DDK antibodies for loading and identification controls. Percent of the MYOC processed band was calculated by densitometry. A) schematic representation of the expression cassettes of the recombinant plasmids. B, C and D: Representative western blots with extracts from transfected cells. B) extracts from HEK293 co-transfected by calcium phosphate. C and D) extracts from primary HTM-137 cells co-transfected by nucleofector electroporation.

    Journal: PLoS ONE

    Article Title: Cystatin A, a Potential Common Link for Mutant Myocilin Causative Glaucoma

    doi: 10.1371/journal.pone.0036301

    Figure Lengend Snippet: Cystatin A inhibits the processing of MYOC wild-type in cultured cells. Recombinant expression plasmids containing tag-fused full coding wild-type MYOC (pMG29), CSTA, and controls plasmids, inactive mutated CSTA (CSTAm) and pEmpty, were generated as indicated in Methods . pMG29 was co-transfected with either pCSTA, pCSTAm or pEmpty (1∶2) and harvested at 48 h post-transfection. Equivalent volumes of cell extracts and of their supernatants were loaded onto 4–15% SDS-PAGE gels, transferred to PVDF membranes and analyzed by immunoblotting. Different MYOC protein forms (full length and processed) were detected with an anti-V5 mouse monoclonal followed by an anti-mouse horseradish peroxidase antibodies. Blots were re-probed with β-actin and DDK antibodies for loading and identification controls. Percent of the MYOC processed band was calculated by densitometry. A) schematic representation of the expression cassettes of the recombinant plasmids. B, C and D: Representative western blots with extracts from transfected cells. B) extracts from HEK293 co-transfected by calcium phosphate. C and D) extracts from primary HTM-137 cells co-transfected by nucleofector electroporation.

    Article Snippet: Aliquots of 4 µl from either the cell lysates or the media were mixed (1∶2 vol) with Laemmli buffer (Bio-Rad, Hercules, CA) containing 5% β-mercaptoethanol and loaded onto 4–15% SDS-PAGE Tris-HCl polyacrylamide gels (Bio-Rad).

    Techniques: Cell Culture, Recombinant, Expressing, Generated, Transfection, SDS Page, Western Blot, Electroporation

    2-DE separation of proteins extracted from leaves of healthy of Las-infected lemon plants. ( A ) Representative leaf and gel containing extracted proteins separated via 2-DE of a healthy lemon plant. ( B ) Representative leaf and gel containing extracted proteins separated via 2-DE of a Las-infected lemon plant. Two-year old healthy plants were either graft-inoculated with side shoots from PCR-confirmed Las-infected bud sticks or uninoculated and leaf samples were analyzed at six months post-inoculation. A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Journal: PLoS ONE

    Article Title: Study on Citrus Response to Huanglongbing Highlights a Down-Regulation of Defense-Related Proteins in Lemon Plants Upon 'Ca. Liberibacter asiaticus' Infection

    doi: 10.1371/journal.pone.0067442

    Figure Lengend Snippet: 2-DE separation of proteins extracted from leaves of healthy of Las-infected lemon plants. ( A ) Representative leaf and gel containing extracted proteins separated via 2-DE of a healthy lemon plant. ( B ) Representative leaf and gel containing extracted proteins separated via 2-DE of a Las-infected lemon plant. Two-year old healthy plants were either graft-inoculated with side shoots from PCR-confirmed Las-infected bud sticks or uninoculated and leaf samples were analyzed at six months post-inoculation. A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Article Snippet: Second dimension electrophoresis was performed in 8-16% gradient SDS-polyacrylamide Tris-HCl gels (Criterion precast gels, Bio-Rad) in a twelve-gel cell system (Criterion Dodeca Cell, Bio-Rad).

    Techniques: Infection, Polymerase Chain Reaction, Stripping Membranes, Staining

    PDQuest-generated master gel image showing the general spot pattern of matched protein spots from the total leaf proteome of healthy or Las-infected lemon plants. Labeled spots were differentially produced in response to Las-infection and described in Table 1 . A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Journal: PLoS ONE

    Article Title: Study on Citrus Response to Huanglongbing Highlights a Down-Regulation of Defense-Related Proteins in Lemon Plants Upon 'Ca. Liberibacter asiaticus' Infection

    doi: 10.1371/journal.pone.0067442

    Figure Lengend Snippet: PDQuest-generated master gel image showing the general spot pattern of matched protein spots from the total leaf proteome of healthy or Las-infected lemon plants. Labeled spots were differentially produced in response to Las-infection and described in Table 1 . A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Article Snippet: Second dimension electrophoresis was performed in 8-16% gradient SDS-polyacrylamide Tris-HCl gels (Criterion precast gels, Bio-Rad) in a twelve-gel cell system (Criterion Dodeca Cell, Bio-Rad).

    Techniques: Generated, Infection, Labeling, Produced, Stripping Membranes, Staining

    Differentially produced protein spots from 2-DE analysis of total leaf proteins from healthy or Las-infected lemon plants. Panels A-M show magnified views of protein spots in representative 2-DE gels containing separated total proteins from leaves of healthy or Las-infected lemon plants. Labeled spots showed significant changes and correspond to the spots presented in in Figure 2 and Tables 2 and 3 . Two-year old healthy plants were either graft-inoculated with side shoots from PCR-confirmed Las-infected bud sticks or uninoculated and leaf samples were analyzed at six months post-inoculation. A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Journal: PLoS ONE

    Article Title: Study on Citrus Response to Huanglongbing Highlights a Down-Regulation of Defense-Related Proteins in Lemon Plants Upon 'Ca. Liberibacter asiaticus' Infection

    doi: 10.1371/journal.pone.0067442

    Figure Lengend Snippet: Differentially produced protein spots from 2-DE analysis of total leaf proteins from healthy or Las-infected lemon plants. Panels A-M show magnified views of protein spots in representative 2-DE gels containing separated total proteins from leaves of healthy or Las-infected lemon plants. Labeled spots showed significant changes and correspond to the spots presented in in Figure 2 and Tables 2 and 3 . Two-year old healthy plants were either graft-inoculated with side shoots from PCR-confirmed Las-infected bud sticks or uninoculated and leaf samples were analyzed at six months post-inoculation. A sum of 200 µg of total protein was separated according to charge on a pH 4-7 IpG strip and according to mass on 8-16% gradient SDS-polyacrylamide Tris-HCl gels. Protein spots were visualized by staining with Coomassie Brilliant Blue (CBB). M r , relative molecular mass; pI, isoelectric point.

    Article Snippet: Second dimension electrophoresis was performed in 8-16% gradient SDS-polyacrylamide Tris-HCl gels (Criterion precast gels, Bio-Rad) in a twelve-gel cell system (Criterion Dodeca Cell, Bio-Rad).

    Techniques: Produced, Infection, Labeling, Polymerase Chain Reaction, Stripping Membranes, Staining

    Subcellular localization of DmLRPPRC2. (A) Fluorescence confocal microscopy images showing HeLa cells ( n = 15) expressing a DmLRPPRC2-GFP fusion protein and counterstained with the mitochondrial marker MitoTracker Deep Red. Scale bar: 10 μm. (B) Subcellular fractionation showing the presence of DmLRPPRC2 in the mitochondrial fraction. Subcellular fractions were separated by SDS-PAGE followed by western blot analysis to detect histone H3 (a nuclear marker), tubulin (a cytosolic marker), VDAC (a mitochondrial marker) and DmLRPPRC2. See also Supplementary Figure S1A and B.

    Journal: Nucleic Acids Research

    Article Title: Drosophila melanogaster LRPPRC2 is involved in coordination of mitochondrial translation

    doi: 10.1093/nar/gku1132

    Figure Lengend Snippet: Subcellular localization of DmLRPPRC2. (A) Fluorescence confocal microscopy images showing HeLa cells ( n = 15) expressing a DmLRPPRC2-GFP fusion protein and counterstained with the mitochondrial marker MitoTracker Deep Red. Scale bar: 10 μm. (B) Subcellular fractionation showing the presence of DmLRPPRC2 in the mitochondrial fraction. Subcellular fractions were separated by SDS-PAGE followed by western blot analysis to detect histone H3 (a nuclear marker), tubulin (a cytosolic marker), VDAC (a mitochondrial marker) and DmLRPPRC2. See also Supplementary Figure S1A and B.

    Article Snippet: All isolated fractions were separated in 4–15% Criterion Tris-HCl precast SDS-PAGE gels (Bio-Rad) and tested by a standard western blotting procedure.

    Techniques: Fluorescence, Confocal Microscopy, Expressing, Marker, Fractionation, SDS Page, Western Blot

    Steady-state levels and activities of the OXPHOS complexes. (A) Western blot analysis of the Lrpprc2 RNAi#1 line (left side) and the Lrpprc2 RNAi#2 line (right side) performed on 20–40 μg whole-body protein extracts from controls and DmLrpprc2 KD third-instar larvae and 6-day-old adults. Protein extracts were separated by standard SDS-PAGE followed by western blot analysis with antibodies against the nuclear-encoded subunit NDUFS3 of complex I, the α-subunit of complex V and VDAC, the latter used as reference for loading. (B) BN-PAGE combined with complex I, complex IV and complex V in-gel activity analyses on the Lrpprc2 RNAi#1 line and the Lrpprc2 RNAi#2 line. BN-PAGE was performed on 75 μg for complex I, 100 μg for complex IV and 150 μg for complex V of mitochondrial protein extracts from control and DmLrpprc2 KD third-instar larvae. The assembly status of complex V (right panel) is black because of color inversion. Loading controls are provided by Coomassie staining to assess the total mitochondrial protein content per sample (only for complex V, left panel) and western blot analysis of VDAC protein levels in mitochondrial protein lysates collected prior to BN-PAGE gel loading. The position of complex I (CI), complex IV (CIV), complex V dimers (CV 2 ), complex V monomers (CV 1 ) and complex V subassembled components ( sub CV a and sub CV b ) are indicated by arrows.

    Journal: Nucleic Acids Research

    Article Title: Drosophila melanogaster LRPPRC2 is involved in coordination of mitochondrial translation

    doi: 10.1093/nar/gku1132

    Figure Lengend Snippet: Steady-state levels and activities of the OXPHOS complexes. (A) Western blot analysis of the Lrpprc2 RNAi#1 line (left side) and the Lrpprc2 RNAi#2 line (right side) performed on 20–40 μg whole-body protein extracts from controls and DmLrpprc2 KD third-instar larvae and 6-day-old adults. Protein extracts were separated by standard SDS-PAGE followed by western blot analysis with antibodies against the nuclear-encoded subunit NDUFS3 of complex I, the α-subunit of complex V and VDAC, the latter used as reference for loading. (B) BN-PAGE combined with complex I, complex IV and complex V in-gel activity analyses on the Lrpprc2 RNAi#1 line and the Lrpprc2 RNAi#2 line. BN-PAGE was performed on 75 μg for complex I, 100 μg for complex IV and 150 μg for complex V of mitochondrial protein extracts from control and DmLrpprc2 KD third-instar larvae. The assembly status of complex V (right panel) is black because of color inversion. Loading controls are provided by Coomassie staining to assess the total mitochondrial protein content per sample (only for complex V, left panel) and western blot analysis of VDAC protein levels in mitochondrial protein lysates collected prior to BN-PAGE gel loading. The position of complex I (CI), complex IV (CIV), complex V dimers (CV 2 ), complex V monomers (CV 1 ) and complex V subassembled components ( sub CV a and sub CV b ) are indicated by arrows.

    Article Snippet: All isolated fractions were separated in 4–15% Criterion Tris-HCl precast SDS-PAGE gels (Bio-Rad) and tested by a standard western blotting procedure.

    Techniques: Western Blot, SDS Page, Polyacrylamide Gel Electrophoresis, Activity Assay, Staining