complete protease inhibitor mixture  (Roche)


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    Structured Review

    Roche complete protease inhibitor mixture
    The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the <t>complete</t> theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,
    Complete Protease Inhibitor Mixture, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete protease inhibitor mixture/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    complete protease inhibitor mixture - by Bioz Stars, 2021-07
    86/100 stars

    Images

    1) Product Images from "Comparative Analysis of the Secretome from a Model Filarial Nematode (Litomosoides sigmodontis) Reveals Maximal Diversity in Gravid Female Parasites *"

    Article Title: Comparative Analysis of the Secretome from a Model Filarial Nematode (Litomosoides sigmodontis) Reveals Maximal Diversity in Gravid Female Parasites *

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1074/mcp.M114.038539

    The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the complete theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,
    Figure Legend Snippet: The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the complete theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,

    Techniques Used: Sequencing

    Pfam enrichment analysis of ESP proteins against the complete theoretical proteome of L. sigmodontis . The fold-enrichment is displayed for each lifecycle stage; DUF290 represents the transthyretin-like protein family.
    Figure Legend Snippet: Pfam enrichment analysis of ESP proteins against the complete theoretical proteome of L. sigmodontis . The fold-enrichment is displayed for each lifecycle stage; DUF290 represents the transthyretin-like protein family.

    Techniques Used: End-sequence Profiling

    Related Articles

    Produced:

    Article Title: Tandem RNA isolation reveals functional rearrangement of RNA-binding proteins on CDKN1B/p27Kip1 3’UTRs in cisplatin treated cells
    Article Snippet: A fragment (580 nts) of the CDS of RASM was amplified as described previously [ ]. .. Biotinylated RNAs were produced with T7-RNA polymerase and the Biotin RNA Labelling Mix (Roche, #11,685,597,910), as instructed by the manufacturer. .. Cell-free extracts were prepared in IP buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 2 mM MgCl2 , 5% glycerol, 1 mM DTT, 0.1% Triton X-100, 0.1 mg/ml heparin, 0.1 mg/ml tRNA, 1 mM phenylmethylsulfonyl chloride (PMSF), 1 µl 20 U/µl RNase OUT (Promega, #10,777,019) and Complete™ mini EDTA-free protease-inhibitor tablets (Roche, #11,836,170,001)) by mechanical disruption of transfected (pEGFPC1-6XHis-FLKSRP) HEK293 cells with glass beads in a Tissue Lyser (Qiagen; 6 × 30 s; 30 Hz, 4°C).

    Protein Extraction:

    Article Title: Differential compartmental processing and phosphorylation of pathogenic human tau and native mouse tau in the line 66 model of frontotemporal dementia
    Article Snippet: .. Protein extraction from brain tissue with and without detergent Five volumes of Tris extraction buffer (30 mm Tris, pH 7.4, containing cOmplete™ protease inhibitor and PhosStop™ phosphatase inhibitor mixture tablets from Roche), with or without the addition of Triton X-100 (0.1% (v/v)), were added to crushed frozen tissue (5 L66 and 3 WT brains pulverized using pestle and mortar with liquid nitrogen), repeatedly pipetted up and down for homogenization and incubated for 30 min at 4 °C. ..

    Protease Inhibitor:

    Article Title: Differential compartmental processing and phosphorylation of pathogenic human tau and native mouse tau in the line 66 model of frontotemporal dementia
    Article Snippet: .. Protein extraction from brain tissue with and without detergent Five volumes of Tris extraction buffer (30 mm Tris, pH 7.4, containing cOmplete™ protease inhibitor and PhosStop™ phosphatase inhibitor mixture tablets from Roche), with or without the addition of Triton X-100 (0.1% (v/v)), were added to crushed frozen tissue (5 L66 and 3 WT brains pulverized using pestle and mortar with liquid nitrogen), repeatedly pipetted up and down for homogenization and incubated for 30 min at 4 °C. ..

    Article Title: Expression and processing of Plasmodium berghei SERA3 during liver stages
    Article Snippet: The resulting PCR products were ligated into the appropriate pGEX vectors and expressed in Escherichia coli BL21 cells (Stratagene) as glutathione S -transferase (GST) fusion proteins. .. GST fusion proteins of PbSERA1 (Leu877 -Gly910 ) or PbSERA3 (Met1 -Phe235 ) were harvested by suspending bacteria expressing SERA proteins in 10 ml buffer A (10 mM EDTA in PBS) in the presence of Complete™ Protease Inhibitor mixture tablets (Roche Molecular Biochemicals) for 30 min followed by sonification. .. The recombinant SERA fusion proteins were purified from the supernatant using glutathione-sepharose as described by the manufacturer (Amersham Biosciences).

    Article Title: Subtle Roles of Down Syndrome Cell Adhesion Molecules in Embryonic Forebrain Development and Neuronal Migration
    Article Snippet: Mouse tail samples (~5 mm) were also collected for DNA extraction and genotyping. .. To verify the absence of DSCAM protein in the DSCAM knockout mouse , protein was extracted from E17.5 brains from knockout and wildtype mice using TRIS-HCL SDS-buffer (65 mM Tris-HCL, 2% SDS) containing cOmplete™ Protease Inhibitor Cocktail (Roche). .. Tissue lysates were cleared by centrifugation and proteins were heat denatured in a mixture of XT sample buffer 4x and XT reducing agent 20x, separated on 4–12% Bis-Tris precast polyacrylamide gels (Criterion XT Bis-Tris Precast Gel, Bio-Rad) in MOPS buffer, and immuno-blotted to nitrocellulose membranes (Trans-Blot Turbo Midi 0.2 μm Nitrocellulose Transfer Packs, Bio-Rad) using a Trans Blot Turbo system (Bio-Rad).

    Article Title: USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity *USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity * S⃞
    Article Snippet: .. Western Blot and Immunoprecipitation —Cell extracts were prepared in lysis buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 1% Nonidet P-40, 1 m m EDTA, 0.5% sodium deoxycholate, and 0.1% SDS) containing 1× protease inhibitor mixture (Complete™; Roche Applied Science). ..

    Article Title: Protein Targets of Inositol Pyrophosphate (5-IP7) in the parasite Trypanosoma cruzi
    Article Snippet: Briefly, beads were washed three times with binding buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Triton X-100), supplemented with 1 mM EDTA or 1 mM MgCl2 . .. Then, total protein lysates from epimastigote and amastigote forms were suspended in lysis buffer [50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Triton X-100, 1X cOmplete™ EDTA-free protease inhibitor, PhosSTOP™ phosphatase inhibitor mixture (Roche) and 1 mM phenylmethanesulfonyl fluoride (PMSF)] supplemented with 1 mM EDTA or 1 mM MgCl2 . ..

    Homogenization:

    Article Title: Differential compartmental processing and phosphorylation of pathogenic human tau and native mouse tau in the line 66 model of frontotemporal dementia
    Article Snippet: .. Protein extraction from brain tissue with and without detergent Five volumes of Tris extraction buffer (30 mm Tris, pH 7.4, containing cOmplete™ protease inhibitor and PhosStop™ phosphatase inhibitor mixture tablets from Roche), with or without the addition of Triton X-100 (0.1% (v/v)), were added to crushed frozen tissue (5 L66 and 3 WT brains pulverized using pestle and mortar with liquid nitrogen), repeatedly pipetted up and down for homogenization and incubated for 30 min at 4 °C. ..

    Incubation:

    Article Title: Differential compartmental processing and phosphorylation of pathogenic human tau and native mouse tau in the line 66 model of frontotemporal dementia
    Article Snippet: .. Protein extraction from brain tissue with and without detergent Five volumes of Tris extraction buffer (30 mm Tris, pH 7.4, containing cOmplete™ protease inhibitor and PhosStop™ phosphatase inhibitor mixture tablets from Roche), with or without the addition of Triton X-100 (0.1% (v/v)), were added to crushed frozen tissue (5 L66 and 3 WT brains pulverized using pestle and mortar with liquid nitrogen), repeatedly pipetted up and down for homogenization and incubated for 30 min at 4 °C. ..

    Expressing:

    Article Title: Expression and processing of Plasmodium berghei SERA3 during liver stages
    Article Snippet: The resulting PCR products were ligated into the appropriate pGEX vectors and expressed in Escherichia coli BL21 cells (Stratagene) as glutathione S -transferase (GST) fusion proteins. .. GST fusion proteins of PbSERA1 (Leu877 -Gly910 ) or PbSERA3 (Met1 -Phe235 ) were harvested by suspending bacteria expressing SERA proteins in 10 ml buffer A (10 mM EDTA in PBS) in the presence of Complete™ Protease Inhibitor mixture tablets (Roche Molecular Biochemicals) for 30 min followed by sonification. .. The recombinant SERA fusion proteins were purified from the supernatant using glutathione-sepharose as described by the manufacturer (Amersham Biosciences).

    Knock-Out:

    Article Title: Subtle Roles of Down Syndrome Cell Adhesion Molecules in Embryonic Forebrain Development and Neuronal Migration
    Article Snippet: Mouse tail samples (~5 mm) were also collected for DNA extraction and genotyping. .. To verify the absence of DSCAM protein in the DSCAM knockout mouse , protein was extracted from E17.5 brains from knockout and wildtype mice using TRIS-HCL SDS-buffer (65 mM Tris-HCL, 2% SDS) containing cOmplete™ Protease Inhibitor Cocktail (Roche). .. Tissue lysates were cleared by centrifugation and proteins were heat denatured in a mixture of XT sample buffer 4x and XT reducing agent 20x, separated on 4–12% Bis-Tris precast polyacrylamide gels (Criterion XT Bis-Tris Precast Gel, Bio-Rad) in MOPS buffer, and immuno-blotted to nitrocellulose membranes (Trans-Blot Turbo Midi 0.2 μm Nitrocellulose Transfer Packs, Bio-Rad) using a Trans Blot Turbo system (Bio-Rad).

    Mouse Assay:

    Article Title: Subtle Roles of Down Syndrome Cell Adhesion Molecules in Embryonic Forebrain Development and Neuronal Migration
    Article Snippet: Mouse tail samples (~5 mm) were also collected for DNA extraction and genotyping. .. To verify the absence of DSCAM protein in the DSCAM knockout mouse , protein was extracted from E17.5 brains from knockout and wildtype mice using TRIS-HCL SDS-buffer (65 mM Tris-HCL, 2% SDS) containing cOmplete™ Protease Inhibitor Cocktail (Roche). .. Tissue lysates were cleared by centrifugation and proteins were heat denatured in a mixture of XT sample buffer 4x and XT reducing agent 20x, separated on 4–12% Bis-Tris precast polyacrylamide gels (Criterion XT Bis-Tris Precast Gel, Bio-Rad) in MOPS buffer, and immuno-blotted to nitrocellulose membranes (Trans-Blot Turbo Midi 0.2 μm Nitrocellulose Transfer Packs, Bio-Rad) using a Trans Blot Turbo system (Bio-Rad).

    Western Blot:

    Article Title: USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity *USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity * S⃞
    Article Snippet: .. Western Blot and Immunoprecipitation —Cell extracts were prepared in lysis buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 1% Nonidet P-40, 1 m m EDTA, 0.5% sodium deoxycholate, and 0.1% SDS) containing 1× protease inhibitor mixture (Complete™; Roche Applied Science). ..

    Immunoprecipitation:

    Article Title: USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity *USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity * S⃞
    Article Snippet: .. Western Blot and Immunoprecipitation —Cell extracts were prepared in lysis buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 1% Nonidet P-40, 1 m m EDTA, 0.5% sodium deoxycholate, and 0.1% SDS) containing 1× protease inhibitor mixture (Complete™; Roche Applied Science). ..

    Article Title: Differential phase partition of a PICS complex is required for piRNA processing and chromosome segregation in C. elegans
    Article Snippet: Primers used for molecular cloning and dual-sgRNA-directed CRISPR/Cas9-mediated gene deletion are listed in Supplementary Table S3. .. Immunoprecipitation followed by mass spectrometry analysis The mix-staged transgenic worms expressing TOFU-6::GFP, TOST-1::GFP, PICS-1::GFP, and ERH-2::GFP were resuspended in the same volume of 2x lysis buffer (50 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol, 1% Triton-X100, Roche®cOmplete™ EDTA-free Protease Inhibitor Cocktail, 10 mM NaF, 2 mM Na3 VO4) and lysed in a FastPrep-24™ 5G Homogenizer. .. The supernatant of lysate was incubated with home-made anti-GFP beads for one hour at 4°C.

    Lysis:

    Article Title: USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity *USP11 Stabilizes HPV-16E7 and Further Modulates the E7 Biological Activity * S⃞
    Article Snippet: .. Western Blot and Immunoprecipitation —Cell extracts were prepared in lysis buffer (50 m m Tris-HCl, pH 8.0, 150 m m NaCl, 1% Nonidet P-40, 1 m m EDTA, 0.5% sodium deoxycholate, and 0.1% SDS) containing 1× protease inhibitor mixture (Complete™; Roche Applied Science). ..

    Article Title: Protein Targets of Inositol Pyrophosphate (5-IP7) in the parasite Trypanosoma cruzi
    Article Snippet: Briefly, beads were washed three times with binding buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.05% Triton X-100), supplemented with 1 mM EDTA or 1 mM MgCl2 . .. Then, total protein lysates from epimastigote and amastigote forms were suspended in lysis buffer [50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1% Triton X-100, 1X cOmplete™ EDTA-free protease inhibitor, PhosSTOP™ phosphatase inhibitor mixture (Roche) and 1 mM phenylmethanesulfonyl fluoride (PMSF)] supplemented with 1 mM EDTA or 1 mM MgCl2 . ..

    Article Title: Differential phase partition of a PICS complex is required for piRNA processing and chromosome segregation in C. elegans
    Article Snippet: Primers used for molecular cloning and dual-sgRNA-directed CRISPR/Cas9-mediated gene deletion are listed in Supplementary Table S3. .. Immunoprecipitation followed by mass spectrometry analysis The mix-staged transgenic worms expressing TOFU-6::GFP, TOST-1::GFP, PICS-1::GFP, and ERH-2::GFP were resuspended in the same volume of 2x lysis buffer (50 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol, 1% Triton-X100, Roche®cOmplete™ EDTA-free Protease Inhibitor Cocktail, 10 mM NaF, 2 mM Na3 VO4) and lysed in a FastPrep-24™ 5G Homogenizer. .. The supernatant of lysate was incubated with home-made anti-GFP beads for one hour at 4°C.

    Mass Spectrometry:

    Article Title: Differential phase partition of a PICS complex is required for piRNA processing and chromosome segregation in C. elegans
    Article Snippet: Primers used for molecular cloning and dual-sgRNA-directed CRISPR/Cas9-mediated gene deletion are listed in Supplementary Table S3. .. Immunoprecipitation followed by mass spectrometry analysis The mix-staged transgenic worms expressing TOFU-6::GFP, TOST-1::GFP, PICS-1::GFP, and ERH-2::GFP were resuspended in the same volume of 2x lysis buffer (50 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol, 1% Triton-X100, Roche®cOmplete™ EDTA-free Protease Inhibitor Cocktail, 10 mM NaF, 2 mM Na3 VO4) and lysed in a FastPrep-24™ 5G Homogenizer. .. The supernatant of lysate was incubated with home-made anti-GFP beads for one hour at 4°C.

    Transgenic Assay:

    Article Title: Differential phase partition of a PICS complex is required for piRNA processing and chromosome segregation in C. elegans
    Article Snippet: Primers used for molecular cloning and dual-sgRNA-directed CRISPR/Cas9-mediated gene deletion are listed in Supplementary Table S3. .. Immunoprecipitation followed by mass spectrometry analysis The mix-staged transgenic worms expressing TOFU-6::GFP, TOST-1::GFP, PICS-1::GFP, and ERH-2::GFP were resuspended in the same volume of 2x lysis buffer (50 mM Tris-HCl pH 8.0, 300 mM NaCl, 10% glycerol, 1% Triton-X100, Roche®cOmplete™ EDTA-free Protease Inhibitor Cocktail, 10 mM NaF, 2 mM Na3 VO4) and lysed in a FastPrep-24™ 5G Homogenizer. .. The supernatant of lysate was incubated with home-made anti-GFP beads for one hour at 4°C.

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    Roche complete protease inhibitor mix
    Soluble JAM-A serum levels do not indicate BBB disturbance in multiple sclerosis and ischemic stroke. A Levels of sJAM-A in serum samples from 45 patients with stable relapsing-remitting MS not receiving secondary prophylactic therapy were measured in duplicates by ELISA and compared to those in 14 untreated MS patients during an acute relapse. B Levels of sJAM-A were measured in serum samples from 13 patients with non-small vessel <t>complete</t> ischemic stroke where a first sample was obtained within the first 3 h of symptom onset and a second sample 24 h after clinical onset. Serum samples were not concentrated before ELISA measurement. Dots represent single values, lines median values. Serum levels in stable and inflammatory-active MS patients were statistically compared by the Mann-Whitney U test, serum levels in stroke patients over time by the Wilcoxon matched pairs test. n.s., not significant.
    Complete Protease Inhibitor Mix, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete protease inhibitor mix/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    complete protease inhibitor mix - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Roche complete protease inhibitor mixture
    The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the <t>complete</t> theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,
    Complete Protease Inhibitor Mixture, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/complete protease inhibitor mixture/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    complete protease inhibitor mixture - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Roche caspase lysis buffer
    Nutlin prevents <t>caspase</t> activation and γH2AX accumulation in response to Wee1 inhibitor and/or gemcitabine A . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, and/or 8μM Nutlin in the absence and presence of 50μM ZVAD-FMK for another 24 hrs. Cells were harvested and immunoblot analysis was performed to detect poly-ADP ribose polymerase (PARP) and γH2AX. B ., C . U2OS cells were treated as in (A). The cells were then fixed and stained for γH2AX by immunofluorescence. Detection and analysis was performed using automated immunofluorescence microscopy (BD Pathway). Figure panel (B) shows images of γH2AX staining for each treatment condition. Quantitation of γH2AX intensities was done using the BD pathway analysis tool and depicted in figure panel (C). Error bars represent the SD, n=3. D . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, 8μM Nutlin in the absence and presence ( Supplementary Figure 1 ) of 50μM ZVAD-FMK for another 24 hrs. The cells were harvested and lysed for caspase activity assay. Fluorescent intensity measurements were obtained for each treatment. The activity (arbitrary units of fluorescence/min) was calculated for each treatment at the linear part of the curve (cf. Supplementary Figure 1 ). Error bars represent the S.D, n=3.
    Caspase Lysis Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/caspase lysis buffer/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    caspase lysis buffer - by Bioz Stars, 2021-07
    86/100 stars
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    86
    Roche applied science complete protease inhibitor mixture tablets
    The spacing of the D3 C X C Cys residues is critical for Sod1 activation. A , mutations were made in the C X C motif of Ccs1 to alter the spacing to either a CC or a C XX C motif without disrupting downstream residues. B , viability tests were performed by plating cells on synthetic <t>complete</t> medium with or without lysine at 30 or 37 °C. C , Sod1 activity for various Ccs1 mutants was quantified; error bars represent S.D. D , the status of the disulfide bond was visualized by lysing cells in the presence of a PEG-maleimide alkylating agent that selectively reacts with free thiols.
    Applied Science Complete Protease Inhibitor Mixture Tablets, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/applied science complete protease inhibitor mixture tablets/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    applied science complete protease inhibitor mixture tablets - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

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    Soluble JAM-A serum levels do not indicate BBB disturbance in multiple sclerosis and ischemic stroke. A Levels of sJAM-A in serum samples from 45 patients with stable relapsing-remitting MS not receiving secondary prophylactic therapy were measured in duplicates by ELISA and compared to those in 14 untreated MS patients during an acute relapse. B Levels of sJAM-A were measured in serum samples from 13 patients with non-small vessel complete ischemic stroke where a first sample was obtained within the first 3 h of symptom onset and a second sample 24 h after clinical onset. Serum samples were not concentrated before ELISA measurement. Dots represent single values, lines median values. Serum levels in stable and inflammatory-active MS patients were statistically compared by the Mann-Whitney U test, serum levels in stroke patients over time by the Wilcoxon matched pairs test. n.s., not significant.

    Journal: PLoS ONE

    Article Title: Evaluation of Soluble Junctional Adhesion Molecule-A as a Biomarker of Human Brain Endothelial Barrier Breakdown

    doi: 10.1371/journal.pone.0013568

    Figure Lengend Snippet: Soluble JAM-A serum levels do not indicate BBB disturbance in multiple sclerosis and ischemic stroke. A Levels of sJAM-A in serum samples from 45 patients with stable relapsing-remitting MS not receiving secondary prophylactic therapy were measured in duplicates by ELISA and compared to those in 14 untreated MS patients during an acute relapse. B Levels of sJAM-A were measured in serum samples from 13 patients with non-small vessel complete ischemic stroke where a first sample was obtained within the first 3 h of symptom onset and a second sample 24 h after clinical onset. Serum samples were not concentrated before ELISA measurement. Dots represent single values, lines median values. Serum levels in stable and inflammatory-active MS patients were statistically compared by the Mann-Whitney U test, serum levels in stroke patients over time by the Wilcoxon matched pairs test. n.s., not significant.

    Article Snippet: Western Blotting For generation of whole cell protein extracts, cells grown to subconfluency in 25 cm2 flasks were stimulated as indicated, washed with icecold PBS and scraped into radioimmunoprecipitation assay (RIPA) buffer composed of 50 mmol/L Tris-HCl, pH 7.4; 150 mmol/L NaCl; 1% Nonidet P40; 0.25% sodium deoxycholate and 1× Roche COMPLETE® protease inhibitor mix (Roche Diagnostics GmbH, Mannheim, Germany).

    Techniques: Mass Spectrometry, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the complete theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Comparative Analysis of the Secretome from a Model Filarial Nematode (Litomosoides sigmodontis) Reveals Maximal Diversity in Gravid Female Parasites *

    doi: 10.1074/mcp.M114.038539

    Figure Lengend Snippet: The ShK domains from L. sigmodontis protein nLs_04059 and its orthologs in other filarial species have a distinct sequence signature. All ShK domains identified in the complete theoretical proteomes of L. sigmodontis, B. malayi, L. loa, W. bancrofti,

    Article Snippet: Soluble WBE was prepared by homogenization in 25 m m ammonium bicarbonate, 1% Rapi Gest SF surfactant (Waters), and cOmplete Protease Inhibitor Mixture (Roche) using a mini-pestle in a microcentrifuge tube.

    Techniques: Sequencing

    Pfam enrichment analysis of ESP proteins against the complete theoretical proteome of L. sigmodontis . The fold-enrichment is displayed for each lifecycle stage; DUF290 represents the transthyretin-like protein family.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Comparative Analysis of the Secretome from a Model Filarial Nematode (Litomosoides sigmodontis) Reveals Maximal Diversity in Gravid Female Parasites *

    doi: 10.1074/mcp.M114.038539

    Figure Lengend Snippet: Pfam enrichment analysis of ESP proteins against the complete theoretical proteome of L. sigmodontis . The fold-enrichment is displayed for each lifecycle stage; DUF290 represents the transthyretin-like protein family.

    Article Snippet: Soluble WBE was prepared by homogenization in 25 m m ammonium bicarbonate, 1% Rapi Gest SF surfactant (Waters), and cOmplete Protease Inhibitor Mixture (Roche) using a mini-pestle in a microcentrifuge tube.

    Techniques: End-sequence Profiling

    Nutlin prevents caspase activation and γH2AX accumulation in response to Wee1 inhibitor and/or gemcitabine A . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, and/or 8μM Nutlin in the absence and presence of 50μM ZVAD-FMK for another 24 hrs. Cells were harvested and immunoblot analysis was performed to detect poly-ADP ribose polymerase (PARP) and γH2AX. B ., C . U2OS cells were treated as in (A). The cells were then fixed and stained for γH2AX by immunofluorescence. Detection and analysis was performed using automated immunofluorescence microscopy (BD Pathway). Figure panel (B) shows images of γH2AX staining for each treatment condition. Quantitation of γH2AX intensities was done using the BD pathway analysis tool and depicted in figure panel (C). Error bars represent the SD, n=3. D . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, 8μM Nutlin in the absence and presence ( Supplementary Figure 1 ) of 50μM ZVAD-FMK for another 24 hrs. The cells were harvested and lysed for caspase activity assay. Fluorescent intensity measurements were obtained for each treatment. The activity (arbitrary units of fluorescence/min) was calculated for each treatment at the linear part of the curve (cf. Supplementary Figure 1 ). Error bars represent the S.D, n=3.

    Journal: Oncotarget

    Article Title: Mdm2 inhibition confers protection of p53-proficient cells from the cytotoxic effects of Wee1 inhibitors

    doi:

    Figure Lengend Snippet: Nutlin prevents caspase activation and γH2AX accumulation in response to Wee1 inhibitor and/or gemcitabine A . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, and/or 8μM Nutlin in the absence and presence of 50μM ZVAD-FMK for another 24 hrs. Cells were harvested and immunoblot analysis was performed to detect poly-ADP ribose polymerase (PARP) and γH2AX. B ., C . U2OS cells were treated as in (A). The cells were then fixed and stained for γH2AX by immunofluorescence. Detection and analysis was performed using automated immunofluorescence microscopy (BD Pathway). Figure panel (B) shows images of γH2AX staining for each treatment condition. Quantitation of γH2AX intensities was done using the BD pathway analysis tool and depicted in figure panel (C). Error bars represent the SD, n=3. D . U2OS cells were treated with 8μM Nutlin for 24 hrs, followed by treatment with 1μM Wee1 inhibitor, 300nM gemcitabine, 8μM Nutlin in the absence and presence ( Supplementary Figure 1 ) of 50μM ZVAD-FMK for another 24 hrs. The cells were harvested and lysed for caspase activity assay. Fluorescent intensity measurements were obtained for each treatment. The activity (arbitrary units of fluorescence/min) was calculated for each treatment at the linear part of the curve (cf. Supplementary Figure 1 ). Error bars represent the S.D, n=3.

    Article Snippet: The pelleted cells were resuspended in 250μl caspase lysis buffer (1M Tris-HCl, 2mM MgCl2 , 150mM NaCl, 10mM DTT, Roche complete mini protease-inhibitor mix).

    Techniques: Activation Assay, Staining, Immunofluorescence, Microscopy, Quantitation Assay, Caspase Activity Assay, Activity Assay, Fluorescence

    The spacing of the D3 C X C Cys residues is critical for Sod1 activation. A , mutations were made in the C X C motif of Ccs1 to alter the spacing to either a CC or a C XX C motif without disrupting downstream residues. B , viability tests were performed by plating cells on synthetic complete medium with or without lysine at 30 or 37 °C. C , Sod1 activity for various Ccs1 mutants was quantified; error bars represent S.D. D , the status of the disulfide bond was visualized by lysing cells in the presence of a PEG-maleimide alkylating agent that selectively reacts with free thiols.

    Journal: The Journal of Biological Chemistry

    Article Title: Copper-zinc superoxide dismutase is activated through a sulfenic acid intermediate at a copper ion entry site

    doi: 10.1074/jbc.M117.775981

    Figure Lengend Snippet: The spacing of the D3 C X C Cys residues is critical for Sod1 activation. A , mutations were made in the C X C motif of Ccs1 to alter the spacing to either a CC or a C XX C motif without disrupting downstream residues. B , viability tests were performed by plating cells on synthetic complete medium with or without lysine at 30 or 37 °C. C , Sod1 activity for various Ccs1 mutants was quantified; error bars represent S.D. D , the status of the disulfide bond was visualized by lysing cells in the presence of a PEG-maleimide alkylating agent that selectively reacts with free thiols.

    Article Snippet: Cells were resuspended in 50 m m NaH2 PO4 , 300 m m NaCl, pH 8.0, with Roche Applied Science cOmplete protease inhibitor mixture tablets.

    Techniques: Activation Assay, Activity Assay