pvdf immobilon p  (Thermo Fisher)


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    Name:
    Tropifluor PVDF Membrane pore size
    Description:
    We have carefully selected and rigorously tested the membrane to provide optimized results in western blotting applications with dioxetane based substrate solutions CSPD or CDP Star substrates • Our Tropifluor PVDF transfer membrane provides the high binding and retention capacity that characterizes PVDF polyvinylidene fluoride membranes • We have carefully selected and rigorously tested the membrane to provide optimized results in western blotting applications with dioxetane based substrate solutions CSPD or CDP Star substrates • PVDF membrane provides high mechanical strength that permits convenient handling and withstands multiple re probing Extend Your Detection LimitsThe low background and resulting superior signal to noise performance obtained with this membrane enables picogram detection levels of proteins in immunoblotting applications We have selected the Tropifluor membrane to yield low background levels with the use of Tropix chemiluminescent substrates and secondary antibody conjugates enabling ultrasensitive detection We recommend this product for all immunoblotting applications where sensitivity is important The membrane has a pore size of 0 45 microns and a binding capacity of 125 µg of protein⁄cm2 Products described herein are guaranteed for 12 months from the date of purchase For Research Use Only Not for use in diagnostics procedures
    Catalog Number:
    t2234
    Price:
    None
    Applications:
    Protein Assays and Analysis|Protein Biology|Western Blot Detection|Western Blotting|Western Blotting Membranes
    Category:
    Gels Fractionation Strips Membranes
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    Structured Review

    Thermo Fisher pvdf immobilon p
    We have carefully selected and rigorously tested the membrane to provide optimized results in western blotting applications with dioxetane based substrate solutions CSPD or CDP Star substrates • Our Tropifluor PVDF transfer membrane provides the high binding and retention capacity that characterizes PVDF polyvinylidene fluoride membranes • We have carefully selected and rigorously tested the membrane to provide optimized results in western blotting applications with dioxetane based substrate solutions CSPD or CDP Star substrates • PVDF membrane provides high mechanical strength that permits convenient handling and withstands multiple re probing Extend Your Detection LimitsThe low background and resulting superior signal to noise performance obtained with this membrane enables picogram detection levels of proteins in immunoblotting applications We have selected the Tropifluor membrane to yield low background levels with the use of Tropix chemiluminescent substrates and secondary antibody conjugates enabling ultrasensitive detection We recommend this product for all immunoblotting applications where sensitivity is important The membrane has a pore size of 0 45 microns and a binding capacity of 125 µg of protein⁄cm2 Products described herein are guaranteed for 12 months from the date of purchase For Research Use Only Not for use in diagnostics procedures
    https://www.bioz.com/result/pvdf immobilon p/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pvdf immobilon p - by Bioz Stars, 2020-11
    99/100 stars

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    Related Articles

    Polyacrylamide Gel Electrophoresis:

    Article Title: Metabolomics profiling reveals new aspects of dolichol biosynthesis in Plasmodium falciparum
    Article Snippet: .. Equal amount of proteins was analyzed using a 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. .. Mouse anti-CPY (1:5,000; Life Technologies, Thermo Fisher Scientific, Waltham, MA, USA) was used as the primary antibody; horseradish peroxidase (HRP)-conjugated anti-mouse antibody (1:5,000 dilution; Thermo Fisher Scientific, Waltham, MA, USA) together with the Supersignal West Femto substrate (Thermo Fisher Scientific, Waltham, MA, USA) were used to detect the target protein.

    Article Title: Diclofenac Inhibits Phorbol Ester-Induced Gene Expression and Production of MUC5AC Mucin via Affecting Degradation of IkBα and Translocation of NF-kB p65 in NCI-H292 Cells
    Article Snippet: .. Detection of proteins by western blot analysisCytosolic, nuclear, and whole cell extracts containing proteins (each 50 µg as proteins) were subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto the polyvinylidene difluoride (PVDF) membrane. ..

    Western Blot:

    Article Title: Strong correlation of ferrochelatase enzymatic activity with Mitoferrin-1 mRNA in lymphoblasts of patients with protoporphyria
    Article Snippet: .. Western blotTotal protein was extracted from cultured cells, 80 μg protein were subjected to SDS-PAGE followed by Western blotting onto PVDF membranes. .. Western transfers were immunoblotted with anti-FECH (1:1000), anti-MFRN1 (1:1000), anti-GAPDH (1:2000) antibodies.

    Article Title: Diclofenac Inhibits Phorbol Ester-Induced Gene Expression and Production of MUC5AC Mucin via Affecting Degradation of IkBα and Translocation of NF-kB p65 in NCI-H292 Cells
    Article Snippet: .. Detection of proteins by western blot analysisCytosolic, nuclear, and whole cell extracts containing proteins (each 50 µg as proteins) were subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto the polyvinylidene difluoride (PVDF) membrane. ..

    Incubation:

    Article Title: Selenium protects against LPS-induced MC3T3-E1 cells apoptosis through modulation of microRNA-155 and PI3K/Akt signaling pathways
    Article Snippet: .. Equal amounts of proteins (20 µg) were separated by 10% SDS-PAGE gels, and then transferred to PVDF membranes, which were blocked for 2 h with 5% non-fat milk before incubated with primary antibodies: Bax(1:400), cyto-C(1:400),p-AKT(1:400) and β-actin (1:1000) overnight at 4 °C. .. The membranes were incubated with HRP-conjugated secondary antibody (Santa Cruz Bio-technology) for 2 h. Finally, the protein bands were visualized using an enhanced chemiluminescence reagent (Pierce).

    SDS Page:

    Article Title: Metabolomics profiling reveals new aspects of dolichol biosynthesis in Plasmodium falciparum
    Article Snippet: .. Equal amount of proteins was analyzed using a 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane. .. Mouse anti-CPY (1:5,000; Life Technologies, Thermo Fisher Scientific, Waltham, MA, USA) was used as the primary antibody; horseradish peroxidase (HRP)-conjugated anti-mouse antibody (1:5,000 dilution; Thermo Fisher Scientific, Waltham, MA, USA) together with the Supersignal West Femto substrate (Thermo Fisher Scientific, Waltham, MA, USA) were used to detect the target protein.

    Article Title: A novel mechanism by which ACTA2-AS1 promotes cervical cancer progression: acting as a ceRNA of miR-143-3p to regulate SMAD3 expression
    Article Snippet: .. Equal amounts of protein samples were separated by 12 percent SDS-PAGE and were transferred into PVDF membranes. .. The membranes were blocked with skim milk for 1 h and then probed with primary antibodies anti‐Smad3 (1:1000, ab28379; Abcam, Cambridge, MA), Bax (1:5000, ab182733; Abcam, Cambridge, MA) and Bcl2 (1:5000, ab117115; Abcam, Cambridge, MA) incubated at 4 °C overnight.

    Article Title: Lack of myosin X enhances osteoclastogenesis and increases cell surface Unc5b in osteoclast-lineage cells
    Article Snippet: .. Protein were resolved by 10% SDS-PAGE gel and transferred to PVDF membranes. .. RNA isolation and Real-Time PCR analysisTotal RNA was isolated from target cells by TRIZOL extraction (Invitrogen).

    Article Title: HIF-1α overexpression in mesenchymal stem cell-derived exosomes mediates cardioprotection in myocardial infarction by enhanced angiogenesis
    Article Snippet: .. Equal quantities of protein were loaded and run on 10% SDS-PAGE gels and then transferred to polyvinylidene difluoride (PVDF) membranes. .. Each membrane was blocked in 5% BSA and subsequently incubated overnight at 4 °C with anti-TSG101 (Abcam, UK) and anti-CD63 (Abcam, UK) antibodies for exosome characterization, anti-HIF-1α antibody (Abcam, UK) for MSCs analysis, and anti-Actin antibody (Beyotime, Shanghai, China) for both.

    Article Title: Selenium protects against LPS-induced MC3T3-E1 cells apoptosis through modulation of microRNA-155 and PI3K/Akt signaling pathways
    Article Snippet: .. Equal amounts of proteins (20 µg) were separated by 10% SDS-PAGE gels, and then transferred to PVDF membranes, which were blocked for 2 h with 5% non-fat milk before incubated with primary antibodies: Bax(1:400), cyto-C(1:400),p-AKT(1:400) and β-actin (1:1000) overnight at 4 °C. .. The membranes were incubated with HRP-conjugated secondary antibody (Santa Cruz Bio-technology) for 2 h. Finally, the protein bands were visualized using an enhanced chemiluminescence reagent (Pierce).

    Article Title: Strong correlation of ferrochelatase enzymatic activity with Mitoferrin-1 mRNA in lymphoblasts of patients with protoporphyria
    Article Snippet: .. Western blotTotal protein was extracted from cultured cells, 80 μg protein were subjected to SDS-PAGE followed by Western blotting onto PVDF membranes. .. Western transfers were immunoblotted with anti-FECH (1:1000), anti-MFRN1 (1:1000), anti-GAPDH (1:2000) antibodies.

    Article Title: Diclofenac Inhibits Phorbol Ester-Induced Gene Expression and Production of MUC5AC Mucin via Affecting Degradation of IkBα and Translocation of NF-kB p65 in NCI-H292 Cells
    Article Snippet: .. Detection of proteins by western blot analysisCytosolic, nuclear, and whole cell extracts containing proteins (each 50 µg as proteins) were subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto the polyvinylidene difluoride (PVDF) membrane. ..

    Cell Culture:

    Article Title: Strong correlation of ferrochelatase enzymatic activity with Mitoferrin-1 mRNA in lymphoblasts of patients with protoporphyria
    Article Snippet: .. Western blotTotal protein was extracted from cultured cells, 80 μg protein were subjected to SDS-PAGE followed by Western blotting onto PVDF membranes. .. Western transfers were immunoblotted with anti-FECH (1:1000), anti-MFRN1 (1:1000), anti-GAPDH (1:2000) antibodies.

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  • 99
    Thermo Fisher pvdf membrane
    Inhibition of RAS stops wt-RAS sequence Philadelphia-like ALL cell growth in the presence of TSLP. (A) MUTZ-5 cells were seeded at 6.5×10 5 /mL density and cultured over 4 days with either 0.5% DMSO (vehicle control), 50 µM Salirasib (indirect Pan-RAS inh.), 10 µM PI-103 (PI3K/mTOR dual inh.), or 5 µM Ruxolitinib (JAK inh.), each in absence or presence of 20 ng/mL human TSLP. Cell count and viability (percentage of acridine orange-positive cells not stained by 4’,6-diamidino-2-phenylindole (DAPI) was determined in a NC-250 automated cell counter daily. The stacked-bar graph on the left side shows the growth rate after the 90 hrs timepoint, averaged from 2 independent experiments, each with triplicate wells. Red error bars are SD from the dead cell fraction while the black error bars show the SD of the viable cells. P -values were calculated in one-way ANOVA from the total cell growth rate and adjusted in a post-hoc Bonferroni multiple comparison. Only relevant P -values are shown in the graph, for a complete list see Supplementary-Tab.S2. (B) The graph shows the cell viability of the experiment in (A) over time. (C) MUTZ-5 cells were pre-treated for 2 hrs with either 0.5% DMSO (vehicle control), 10 µM PI-103 (PI3K/mTOR dual inh.), 50 µM Salirasib (indirect Pan-RAS inh.), 5 µM Ruxolitinib (JAK inh.), 50 µM Vemurafenib (Pan-Raf inh.), or 25 µM II-B08 (PTPN11 inh.), and then stimulated with 20 ng/mL human TSLP for 10 min followed by cell lysis. Each lysate sample was split up for analysis in RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down (left) and lysate samples (right) were loaded on separate gels. An <t>SDS-PAGE</t> followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same <t>PVDF-membrane,</t> membranes were stripped and reprobed with new antibodies. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band.
    Pvdf Membrane, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1567 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvdf membrane/product/Thermo Fisher
    Average 99 stars, based on 1567 article reviews
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    pvdf membrane - by Bioz Stars, 2020-11
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    94
    Thermo Fisher pvdf
    ( A ) Transmission electron microscopy (TEM) image of pristine carbon nanotubes <t>(CNTs)</t> dispersed in the water; ( B ) TEM image of ionic liquids (ILs) coated CNTs dispersed in the water; ( C ) Thermogravimetric analysis (TGA) curves of pristine CNTs, pure ILs and ILs modified CNTs, respectively; ( D ) Raman spectra of pristine CNTs and IL coated CNTs (i.e., CNTs/IL = 1/10); ( E ) TEM image of <t>PVDF/CNTs</t> (100/1) composite; ( F ) TEM image of PVDF/ IL-CNTs (100/10-1) nanocomposites.
    Pvdf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvdf/product/Thermo Fisher
    Average 94 stars, based on 119 article reviews
    Price from $9.99 to $1999.99
    pvdf - by Bioz Stars, 2020-11
    94/100 stars
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    99
    Thermo Fisher pvdf pre cut blotting membrane
    Top Panel: <t>SDS-PAGE</t> analysis of time course digestion of Vip3Ab1 and Vip3Bc1 with H. zea and P. includens gut enzymes. Vip3Ab1 and Vip3Bc1 proteins (150 µg/mL) were incubated with gut fluids from H. zea (left) and P. includens (right) at 30 °C for various time intervals at pH 10.0. Bottom Panel: SDS-PAGE analysis of overnight digestion of Vip3 chimeras with H. zea gut enzymes. Vip3_AB and Vip3_BA proteins (110 µg/ml) were incubated with H. zea gut fluids for 16 hours at 30 °C in a total volume of 100 µL at pH 10.0. All reactions were stopped with protease inhibitors and 30 µL of the reaction loaded as described in Materials and Methods. Equivalent lanes were loaded and blotted onto a <t>PVDF</t> membrane for N-terminal sequencing.
    Pvdf Pre Cut Blotting Membrane, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pvdf pre cut blotting membrane/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pvdf pre cut blotting membrane - by Bioz Stars, 2020-11
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    Inhibition of RAS stops wt-RAS sequence Philadelphia-like ALL cell growth in the presence of TSLP. (A) MUTZ-5 cells were seeded at 6.5×10 5 /mL density and cultured over 4 days with either 0.5% DMSO (vehicle control), 50 µM Salirasib (indirect Pan-RAS inh.), 10 µM PI-103 (PI3K/mTOR dual inh.), or 5 µM Ruxolitinib (JAK inh.), each in absence or presence of 20 ng/mL human TSLP. Cell count and viability (percentage of acridine orange-positive cells not stained by 4’,6-diamidino-2-phenylindole (DAPI) was determined in a NC-250 automated cell counter daily. The stacked-bar graph on the left side shows the growth rate after the 90 hrs timepoint, averaged from 2 independent experiments, each with triplicate wells. Red error bars are SD from the dead cell fraction while the black error bars show the SD of the viable cells. P -values were calculated in one-way ANOVA from the total cell growth rate and adjusted in a post-hoc Bonferroni multiple comparison. Only relevant P -values are shown in the graph, for a complete list see Supplementary-Tab.S2. (B) The graph shows the cell viability of the experiment in (A) over time. (C) MUTZ-5 cells were pre-treated for 2 hrs with either 0.5% DMSO (vehicle control), 10 µM PI-103 (PI3K/mTOR dual inh.), 50 µM Salirasib (indirect Pan-RAS inh.), 5 µM Ruxolitinib (JAK inh.), 50 µM Vemurafenib (Pan-Raf inh.), or 25 µM II-B08 (PTPN11 inh.), and then stimulated with 20 ng/mL human TSLP for 10 min followed by cell lysis. Each lysate sample was split up for analysis in RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down (left) and lysate samples (right) were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band.

    Journal: bioRxiv

    Article Title: RAS activation via CRLF2 signaling is a widespread mechanism in Down syndrome acute lymphoblastic leukemia regardless of RAS mutations

    doi: 10.1101/2020.02.03.931725

    Figure Lengend Snippet: Inhibition of RAS stops wt-RAS sequence Philadelphia-like ALL cell growth in the presence of TSLP. (A) MUTZ-5 cells were seeded at 6.5×10 5 /mL density and cultured over 4 days with either 0.5% DMSO (vehicle control), 50 µM Salirasib (indirect Pan-RAS inh.), 10 µM PI-103 (PI3K/mTOR dual inh.), or 5 µM Ruxolitinib (JAK inh.), each in absence or presence of 20 ng/mL human TSLP. Cell count and viability (percentage of acridine orange-positive cells not stained by 4’,6-diamidino-2-phenylindole (DAPI) was determined in a NC-250 automated cell counter daily. The stacked-bar graph on the left side shows the growth rate after the 90 hrs timepoint, averaged from 2 independent experiments, each with triplicate wells. Red error bars are SD from the dead cell fraction while the black error bars show the SD of the viable cells. P -values were calculated in one-way ANOVA from the total cell growth rate and adjusted in a post-hoc Bonferroni multiple comparison. Only relevant P -values are shown in the graph, for a complete list see Supplementary-Tab.S2. (B) The graph shows the cell viability of the experiment in (A) over time. (C) MUTZ-5 cells were pre-treated for 2 hrs with either 0.5% DMSO (vehicle control), 10 µM PI-103 (PI3K/mTOR dual inh.), 50 µM Salirasib (indirect Pan-RAS inh.), 5 µM Ruxolitinib (JAK inh.), 50 µM Vemurafenib (Pan-Raf inh.), or 25 µM II-B08 (PTPN11 inh.), and then stimulated with 20 ng/mL human TSLP for 10 min followed by cell lysis. Each lysate sample was split up for analysis in RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down (left) and lysate samples (right) were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band.

    Article Snippet: SDS-PAGE with 11%-resolving/5%-stacking acrylamide gels, and WB on PVDF-membrane (Cat.#88518; ThermoFisher Scientific) were performed according to the standard protocol of the equipment-manufacturer (Bio-Rad Laboratories).

    Techniques: Inhibition, Sequencing, Cell Culture, Cell Counting, Staining, Lysis, Pull Down Assay, SDS Page, Western Blot, Labeling

    Human Ph-like B-ALL (spontaneous CRLF2-rearrangment and JAK2R683G mutation) cells activate wildtype RAS and RAS-interacting proteins upon TSLP induction. MUTZ-5 cells (Human Ph-like B-ALL cells bearing CRLF2-rearranged and spontaneous JAK2R683G mutation) were stimulated with 20 ng/mL human TSLP (maximal effective TSLP-concentration, Supplementary-Fig.S3B) for 10 min before cell lysis. Each cell lysate was split up for analysis in RAS-GTP pull-down assay and for total protein signal. (A) RAS-GTP pull-down and lysate samples were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, each membrane part was stripped and reprobed with new antibodies. RAS-GTP pull-down samples are on the left side while the right-hand side blots show whole cell lysates of the same samples. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band. The grey arrow shows the unspecific signal of the GST-RAS binding domain (RBD) used in the active RAS pull-down assay acting as a loading control. The experiment was repeated 5 times independently and the graphs show the quantification for active RAS (RAS-GTP), phosphorylated MEK1/2 (phospho-MEK1/2), JAK2 (phospho-JAK2), and PTPN11 (phosho-PTPN11). Beta-actin and total protein signals were used as a loading control to normalize samples. (B) A blot separate from (A) demonstrates the TSLP-inducibility of RAS-effector bRAF. (C) RAS-GTP quantification of 5 independent ELISA experiments in which RAS activity of TSLP-induced MUTZ-5 cells was measured using a different, ELISA-specific active RAS pull-down assay. (D) TSLP-induced MUTZ-5 cells were probed for the presence of activated isoforms KRAS-GTP, HRAS-GTP, or NRAS-GTP (blots on the left). The blots on the right show the total expression of the respective RAS proteins and the graphs show the signal fold change over uninduced MUTZ-5 cells for KRAS-GTP, HRAS-GTP and NRAS-GTP of 4 independent experiments. All error bars are SD; P -values were calculated using Student’s T-test and are adjusted with a Bonferroni-correction for sequential multiple comparison.

    Journal: bioRxiv

    Article Title: RAS activation via CRLF2 signaling is a widespread mechanism in Down syndrome acute lymphoblastic leukemia regardless of RAS mutations

    doi: 10.1101/2020.02.03.931725

    Figure Lengend Snippet: Human Ph-like B-ALL (spontaneous CRLF2-rearrangment and JAK2R683G mutation) cells activate wildtype RAS and RAS-interacting proteins upon TSLP induction. MUTZ-5 cells (Human Ph-like B-ALL cells bearing CRLF2-rearranged and spontaneous JAK2R683G mutation) were stimulated with 20 ng/mL human TSLP (maximal effective TSLP-concentration, Supplementary-Fig.S3B) for 10 min before cell lysis. Each cell lysate was split up for analysis in RAS-GTP pull-down assay and for total protein signal. (A) RAS-GTP pull-down and lysate samples were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, each membrane part was stripped and reprobed with new antibodies. RAS-GTP pull-down samples are on the left side while the right-hand side blots show whole cell lysates of the same samples. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band. The grey arrow shows the unspecific signal of the GST-RAS binding domain (RBD) used in the active RAS pull-down assay acting as a loading control. The experiment was repeated 5 times independently and the graphs show the quantification for active RAS (RAS-GTP), phosphorylated MEK1/2 (phospho-MEK1/2), JAK2 (phospho-JAK2), and PTPN11 (phosho-PTPN11). Beta-actin and total protein signals were used as a loading control to normalize samples. (B) A blot separate from (A) demonstrates the TSLP-inducibility of RAS-effector bRAF. (C) RAS-GTP quantification of 5 independent ELISA experiments in which RAS activity of TSLP-induced MUTZ-5 cells was measured using a different, ELISA-specific active RAS pull-down assay. (D) TSLP-induced MUTZ-5 cells were probed for the presence of activated isoforms KRAS-GTP, HRAS-GTP, or NRAS-GTP (blots on the left). The blots on the right show the total expression of the respective RAS proteins and the graphs show the signal fold change over uninduced MUTZ-5 cells for KRAS-GTP, HRAS-GTP and NRAS-GTP of 4 independent experiments. All error bars are SD; P -values were calculated using Student’s T-test and are adjusted with a Bonferroni-correction for sequential multiple comparison.

    Article Snippet: SDS-PAGE with 11%-resolving/5%-stacking acrylamide gels, and WB on PVDF-membrane (Cat.#88518; ThermoFisher Scientific) were performed according to the standard protocol of the equipment-manufacturer (Bio-Rad Laboratories).

    Techniques: Mutagenesis, Concentration Assay, Lysis, Pull Down Assay, SDS Page, Western Blot, Labeling, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Expressing

    Direct wtRAS activation can precede PI3K/mTOR pathway activation, and the resulting PI3K downstream signaling activity can be blocked by RAS inhibitor. (A) Effect of TSLP induction over time. MUTZ-5 cells were incubated with 20 ng/mL human TSLP at 37 °C and 5% CO 2 for the indicated time points (0 min to 18 hrs) before cell lysis. Due to the centrifugation step of the suspension cells the TSLP is able to act for 5 min before lysis at timepoint 0. Each cell lysate was split up for analysis in RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down and lysate samples were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. RAS-GTP pull-down elutions are on the left side while the right-hand side blots show whole cell lysates of the same samples. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band. (B) Activation of PI3K/mTOR downstream target rpS6 protein was monitored via PLA in high-throughput microscopy. MUTZ-5 cells were either not induced or induced with 20 ng/mL TSLP for 10 min. Where indicated, cells were pre-treated for 3 hrs with either DMSO (vehicle control), RAS inhibitor, or JAK inhibitor. Cells were fixed and permeabilized in a 96 well plate. After blocking, antibodies against phosphorylated rpS6 and total rpS6 were used in conjunction with PLA rabbit and mouse probes to allow specific readout of rpS6 activation in single cells in a high-throughput manner. Histograms show the distribution for a single experiment of the number of PLA spots in cells with at least 1 PLA spot (assay control is only shown in the bar graph). A minimum of 600 cells were analyzed per sample. Non-linear Gaussian fitting curves were plotted. Fluorescent microscope images show examples of PLA spots in MUTZ-5 cells for the respective treatment; white scale bars are 20 µm long. (C) The bar graph summarizes the average PLA spot counts of 3 independent experiments. Error bars are SD and P -values were determined in one-way ANOVA and post-hoc Bonferroni multiple comparison.

    Journal: bioRxiv

    Article Title: RAS activation via CRLF2 signaling is a widespread mechanism in Down syndrome acute lymphoblastic leukemia regardless of RAS mutations

    doi: 10.1101/2020.02.03.931725

    Figure Lengend Snippet: Direct wtRAS activation can precede PI3K/mTOR pathway activation, and the resulting PI3K downstream signaling activity can be blocked by RAS inhibitor. (A) Effect of TSLP induction over time. MUTZ-5 cells were incubated with 20 ng/mL human TSLP at 37 °C and 5% CO 2 for the indicated time points (0 min to 18 hrs) before cell lysis. Due to the centrifugation step of the suspension cells the TSLP is able to act for 5 min before lysis at timepoint 0. Each cell lysate was split up for analysis in RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down and lysate samples were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. RAS-GTP pull-down elutions are on the left side while the right-hand side blots show whole cell lysates of the same samples. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band. (B) Activation of PI3K/mTOR downstream target rpS6 protein was monitored via PLA in high-throughput microscopy. MUTZ-5 cells were either not induced or induced with 20 ng/mL TSLP for 10 min. Where indicated, cells were pre-treated for 3 hrs with either DMSO (vehicle control), RAS inhibitor, or JAK inhibitor. Cells were fixed and permeabilized in a 96 well plate. After blocking, antibodies against phosphorylated rpS6 and total rpS6 were used in conjunction with PLA rabbit and mouse probes to allow specific readout of rpS6 activation in single cells in a high-throughput manner. Histograms show the distribution for a single experiment of the number of PLA spots in cells with at least 1 PLA spot (assay control is only shown in the bar graph). A minimum of 600 cells were analyzed per sample. Non-linear Gaussian fitting curves were plotted. Fluorescent microscope images show examples of PLA spots in MUTZ-5 cells for the respective treatment; white scale bars are 20 µm long. (C) The bar graph summarizes the average PLA spot counts of 3 independent experiments. Error bars are SD and P -values were determined in one-way ANOVA and post-hoc Bonferroni multiple comparison.

    Article Snippet: SDS-PAGE with 11%-resolving/5%-stacking acrylamide gels, and WB on PVDF-membrane (Cat.#88518; ThermoFisher Scientific) were performed according to the standard protocol of the equipment-manufacturer (Bio-Rad Laboratories).

    Techniques: Activation Assay, Activity Assay, Incubation, Lysis, Centrifugation, Pull Down Assay, SDS Page, Western Blot, Labeling, Proximity Ligation Assay, High Throughput Screening Assay, Microscopy, Blocking Assay, Spot Test

    70% of primary bone marrow presentation samples of Down syndrome ALL patients show activated and/or TSLP-inducible RAS, regardless of mutations status. Primary presentation samples of DS-ALL patients were cultured for 2 days (see Supplementary-Fig.S4A legend for details) and then induced for 10 min with 20 ng/mL TSLP (or not induced) in serum-reduced medium. Lysates were analyzed for RAS activity in WB pull-down (A) or ELISA (B) using an ELISA-specific RAS pull-down assay. (A) Each lysate was split up for analysis in Western blot RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down (left) and lysate samples (right) were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band; the grey arrow shows the loading of the GST-RBD in the pull-down assay. (B) The RAS activity pattern in the patient samples from (A) was confirmed via ELISA measurement of RAS-activity in aliquots that were independently thawed and processed as described above The line graph illustrates the four main patterns observed for RAS activity in primary ALL patient samples. (C) Shows an overview of the ELISA-measured RAS activity for the DS-ALL cohort at diagnosis (not enough cell material was available for DS26, DS29 and DS30). The RAS-GTP pull-down for ELISA was performed on lysates from cells at minimum 75% viability at a 100 ng/μL total protein concentration. In parallel, uninduced MUTZ-5 cells were subjected to the same treatment as the primary patient cells and were used to normalize all RAS activities. Brackets on top indicate the groups of the four RAS activity patterns presented in (A, B). For visualization purposes only in this graph, basal RAS activity over 50% of MUTZ-5 basal RAS activity was grouped as high RAS activity while an increase by at least 10% RAS-GTP in TSLP-stimulated samples over uninduced samples in ELISA was classed as TSLP-inducible RAS. For visualization purposes only in this graph, phosphorylation levels measured in WB for JAK2 were categorized as –(negative) = 0.00-0.05; + = 0.05-0.50; ++ = 0.50-1.00; +++ =1.00-2.00, and CRLF2 protein levels were categorized as –(negative) = 0.00-0.05; + = 0.05-0.20; ++ = 0.20-0.50; +++ =0.50-1.50. None of the arbitrary threshold groupings defined above were used in any of the PCA or clustering analysis shown later ( Fig.3 , Supplementary-FigS5). Known CRLF2-rearrangements are marked (R). All values are normalized to those measured for uninduced MUTZ-5 cells.). Outcome of leukemia is given (white = good outcome, black = poor outcome), and the presence of RAS mutations (blue) or JAK2 mutations (red) are specified (grey means unsequenced samples). The groups at the right end of the bar graph (separated by the black bar) show average RAS activities for patient/sample groups other than DS-ALL-diagnosis: Non-DS (NDS) at presentation, DS complete remission (CR) and DS/NDS at relapse. For an overview of the complete Western blot data and the quantified activities and protein expression of STAT5, JAK2, MEK1/2, ERK1/2 and rpS6 of all individual samples, see Supplementary-Fig.S4.

    Journal: bioRxiv

    Article Title: RAS activation via CRLF2 signaling is a widespread mechanism in Down syndrome acute lymphoblastic leukemia regardless of RAS mutations

    doi: 10.1101/2020.02.03.931725

    Figure Lengend Snippet: 70% of primary bone marrow presentation samples of Down syndrome ALL patients show activated and/or TSLP-inducible RAS, regardless of mutations status. Primary presentation samples of DS-ALL patients were cultured for 2 days (see Supplementary-Fig.S4A legend for details) and then induced for 10 min with 20 ng/mL TSLP (or not induced) in serum-reduced medium. Lysates were analyzed for RAS activity in WB pull-down (A) or ELISA (B) using an ELISA-specific RAS pull-down assay. (A) Each lysate was split up for analysis in Western blot RAS-GTP pull-down assay and for total protein signal. RAS-GTP pull-down (left) and lysate samples (right) were loaded on separate gels. An SDS-PAGE followed by Western blotting was performed. To assess the total protein and phosphorylated protein amounts on the same PVDF-membrane, membranes were stripped and reprobed with new antibodies. Antibody-targets are labeled on the right side of each image with black arrows indicating the respective protein band; the grey arrow shows the loading of the GST-RBD in the pull-down assay. (B) The RAS activity pattern in the patient samples from (A) was confirmed via ELISA measurement of RAS-activity in aliquots that were independently thawed and processed as described above The line graph illustrates the four main patterns observed for RAS activity in primary ALL patient samples. (C) Shows an overview of the ELISA-measured RAS activity for the DS-ALL cohort at diagnosis (not enough cell material was available for DS26, DS29 and DS30). The RAS-GTP pull-down for ELISA was performed on lysates from cells at minimum 75% viability at a 100 ng/μL total protein concentration. In parallel, uninduced MUTZ-5 cells were subjected to the same treatment as the primary patient cells and were used to normalize all RAS activities. Brackets on top indicate the groups of the four RAS activity patterns presented in (A, B). For visualization purposes only in this graph, basal RAS activity over 50% of MUTZ-5 basal RAS activity was grouped as high RAS activity while an increase by at least 10% RAS-GTP in TSLP-stimulated samples over uninduced samples in ELISA was classed as TSLP-inducible RAS. For visualization purposes only in this graph, phosphorylation levels measured in WB for JAK2 were categorized as –(negative) = 0.00-0.05; + = 0.05-0.50; ++ = 0.50-1.00; +++ =1.00-2.00, and CRLF2 protein levels were categorized as –(negative) = 0.00-0.05; + = 0.05-0.20; ++ = 0.20-0.50; +++ =0.50-1.50. None of the arbitrary threshold groupings defined above were used in any of the PCA or clustering analysis shown later ( Fig.3 , Supplementary-FigS5). Known CRLF2-rearrangements are marked (R). All values are normalized to those measured for uninduced MUTZ-5 cells.). Outcome of leukemia is given (white = good outcome, black = poor outcome), and the presence of RAS mutations (blue) or JAK2 mutations (red) are specified (grey means unsequenced samples). The groups at the right end of the bar graph (separated by the black bar) show average RAS activities for patient/sample groups other than DS-ALL-diagnosis: Non-DS (NDS) at presentation, DS complete remission (CR) and DS/NDS at relapse. For an overview of the complete Western blot data and the quantified activities and protein expression of STAT5, JAK2, MEK1/2, ERK1/2 and rpS6 of all individual samples, see Supplementary-Fig.S4.

    Article Snippet: SDS-PAGE with 11%-resolving/5%-stacking acrylamide gels, and WB on PVDF-membrane (Cat.#88518; ThermoFisher Scientific) were performed according to the standard protocol of the equipment-manufacturer (Bio-Rad Laboratories).

    Techniques: Cell Culture, Activity Assay, Western Blot, Enzyme-linked Immunosorbent Assay, Pull Down Assay, SDS Page, Labeling, Protein Concentration, Expressing

    ( A ) Transmission electron microscopy (TEM) image of pristine carbon nanotubes (CNTs) dispersed in the water; ( B ) TEM image of ionic liquids (ILs) coated CNTs dispersed in the water; ( C ) Thermogravimetric analysis (TGA) curves of pristine CNTs, pure ILs and ILs modified CNTs, respectively; ( D ) Raman spectra of pristine CNTs and IL coated CNTs (i.e., CNTs/IL = 1/10); ( E ) TEM image of PVDF/CNTs (100/1) composite; ( F ) TEM image of PVDF/ IL-CNTs (100/10-1) nanocomposites.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: ( A ) Transmission electron microscopy (TEM) image of pristine carbon nanotubes (CNTs) dispersed in the water; ( B ) TEM image of ionic liquids (ILs) coated CNTs dispersed in the water; ( C ) Thermogravimetric analysis (TGA) curves of pristine CNTs, pure ILs and ILs modified CNTs, respectively; ( D ) Raman spectra of pristine CNTs and IL coated CNTs (i.e., CNTs/IL = 1/10); ( E ) TEM image of PVDF/CNTs (100/1) composite; ( F ) TEM image of PVDF/ IL-CNTs (100/10-1) nanocomposites.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Modification

    Frequency dependency of dielectric constant ( A , C ) and loss tangent ( B , D ) of samples including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), as-irradiated PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs with various CNTs contents.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Frequency dependency of dielectric constant ( A , C ) and loss tangent ( B , D ) of samples including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), as-irradiated PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs with various CNTs contents.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Irradiation

    Raman spectra of pristine CNTs, IL coated CNTs (CNTs/IL = 1/10), PVDF/IL-CNTs (100/10-1), as-irradiated PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1) samples, respectively.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Raman spectra of pristine CNTs, IL coated CNTs (CNTs/IL = 1/10), PVDF/IL-CNTs (100/10-1), as-irradiated PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1) samples, respectively.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Irradiation

    TEM and SEM images of nano-PVDF/IL-CNTs samples with various content of CNTs: ( A ) and ( a ): 100/10-0.1; ( B ) and ( b ): 100/10-0.5; ( C ) and ( c ): 100/10-1; ( D ) and ( d ) 100/10-2, respectively.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: TEM and SEM images of nano-PVDF/IL-CNTs samples with various content of CNTs: ( A ) and ( a ): 100/10-0.1; ( B ) and ( b ): 100/10-0.5; ( C ) and ( c ): 100/10-1; ( D ) and ( d ) 100/10-2, respectively.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Transmission Electron Microscopy

    TEM image of as-irradiated poly(vinylidene fluoride) (PVDF)/IL-CNTs (100/10-1).

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: TEM image of as-irradiated poly(vinylidene fluoride) (PVDF)/IL-CNTs (100/10-1).

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Transmission Electron Microscopy, Irradiation

    Schematic diagrams for the formation of ionic nanoclusters and CNTs structures in nano-PVDF/IL-CNTs composites. ( A ) First grounding of CNTs with IL resulted in IL-layer-coated CNTs; ( B ) melt-blending PVDF with such IL-layer-coated CNTs fabricated PVDF/IL-CNTs nanocomposites with homogeneously dispersed CNTs; ( C ) The PVDF/IL-CNTs nanocomposites then exposed upon electron beam irradiated (EBI) at room temperature in the air, whereas IL molecules in-suit grafted onto chains of PVDF; ( D ) the as-irradiated PVDF/IL-CNTs samples were heated to 210 °C and held there for 30 min, and microphase separation of IL grafted PVDF (PVDF- g -IL) chains occurred in the melt. A following cooling procedure led to the nano-PVDF/IL-CNTs composites with PVDF- g -IL nanodomains (i.e., IL nanoclusters).

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Schematic diagrams for the formation of ionic nanoclusters and CNTs structures in nano-PVDF/IL-CNTs composites. ( A ) First grounding of CNTs with IL resulted in IL-layer-coated CNTs; ( B ) melt-blending PVDF with such IL-layer-coated CNTs fabricated PVDF/IL-CNTs nanocomposites with homogeneously dispersed CNTs; ( C ) The PVDF/IL-CNTs nanocomposites then exposed upon electron beam irradiated (EBI) at room temperature in the air, whereas IL molecules in-suit grafted onto chains of PVDF; ( D ) the as-irradiated PVDF/IL-CNTs samples were heated to 210 °C and held there for 30 min, and microphase separation of IL grafted PVDF (PVDF- g -IL) chains occurred in the melt. A following cooling procedure led to the nano-PVDF/IL-CNTs composites with PVDF- g -IL nanodomains (i.e., IL nanoclusters).

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Irradiation

    Strain-stress curves of neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), ir-PVDF/IL-CNTs (100/10-1), and nano-PVDF/IL-CNTs (100/10-1).

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Strain-stress curves of neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), ir-PVDF/IL-CNTs (100/10-1), and nano-PVDF/IL-CNTs (100/10-1).

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques:

    Characterization of crystallization behaviors of nano-PVDF/IL-CNTs (100/10-1) nanocomposite in comparison with its counterparts, including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1) and ir-PVDF/IL-CNTs (100/10-1). ( A ): Differential scanning calorimeter (DSC) cooling curves with a 10 °C /min cooling rate; ( B ) Wide-angle X-ray diffraction (WAXD) patterns in the range of 10–40° with a scanning rate of 1°/min.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Characterization of crystallization behaviors of nano-PVDF/IL-CNTs (100/10-1) nanocomposite in comparison with its counterparts, including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1) and ir-PVDF/IL-CNTs (100/10-1). ( A ): Differential scanning calorimeter (DSC) cooling curves with a 10 °C /min cooling rate; ( B ) Wide-angle X-ray diffraction (WAXD) patterns in the range of 10–40° with a scanning rate of 1°/min.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Crystallization Assay

    ( A ) TEM image of nano-PVDF/IL-CNTs (100/10-1) and ( B ) Small-angle X-ray scattering (SAXS) patterns of PVDF/IL-CNTs (100/10-1), irradiated-PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1) samples.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: ( A ) TEM image of nano-PVDF/IL-CNTs (100/10-1) and ( B ) Small-angle X-ray scattering (SAXS) patterns of PVDF/IL-CNTs (100/10-1), irradiated-PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1) samples.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Transmission Electron Microscopy, Irradiation

    Electrical properties of PVDF and PVDF-based samples. ( A , B ) are AC conductivity and surface resistivity (Rs) of typical samples including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), ir-PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1), respectively; ( C ) AC conductivity of nano-PVDF/IL-CNTs samples with various CNTs loading levels; ( D ) Rv values of three typical systems, including PVDF/IL-CNTs, as-irradiated PVDF/IL-CNTs and nano-PVDF/IL-CNTs with different CNTs contents.

    Journal: Polymers

    Article Title: Towards Flexible Dielectric Materials with High Dielectric Constant and Low Loss: PVDF Nanocomposites with both Homogenously Dispersed CNTs and Ionic Liquids Nanodomains

    doi: 10.3390/polym9110562

    Figure Lengend Snippet: Electrical properties of PVDF and PVDF-based samples. ( A , B ) are AC conductivity and surface resistivity (Rs) of typical samples including neat PVDF, PVDF/CNTs (100/1), PVDF/IL-CNTs (100/10-1), ir-PVDF/IL-CNTs (100/10-1) and nano-PVDF/IL-CNTs (100/10-1), respectively; ( C ) AC conductivity of nano-PVDF/IL-CNTs samples with various CNTs loading levels; ( D ) Rv values of three typical systems, including PVDF/IL-CNTs, as-irradiated PVDF/IL-CNTs and nano-PVDF/IL-CNTs with different CNTs contents.

    Article Snippet: Preparation of PVDF/MWCNTs Nanocomposites with IL Nanodomains The final nanocomposites were prepared by the following steps: (1) preparation of ILs modified CNTs (IL/CNTs)—the MWCNTs were first ground with the ILs at room temperature with various weight ratios, MWCNTs bulky gel was thus prepared and termed IL/CNTs; (2) preparation of PVDF/IL-CNTs nanocomposites—the IL/CNTs were melt compounded with PVDF at 190 °C using a Haake mixer (Haake Polylab QC), (Thermo Fisher Scientific, Waltham, MA, USA) with the screw rotation speed of 50 rpm, the PVDF/IL-CNTs blends were then hot-pressed at 190 °C into films with the thickness of about 300 μm; (3) preparation of EB irradiated PVDF/IL-CNTs nanocomposites—the hot pressed films were irradiated at a dose of 45 kGy in air at room temperature using an electron beam accelerator, the as-irradiated PVDF/IL-CNTs films were thus fabricated and termed ir-PVDF/IL-CNTs (the acceleration energy and beam current were 2.5 MeV and 17 mA, respectively); and (4) preparation of PVDF/CNTs with IL nanodomains—the EBI irradiated PVDF/IL-CNTs nanocomposites were heated to 210 °C for 30 min, followed by a cooling procedure, and the PVDF nanocomposites with IL nanodomains were then prepared; they were termed nano-PVDF/IL-CNTs nanocomposites (for instance, the sample of PVDF/IL-CNTs 100/10-1 meant that the weight ratios of PVDF, IL and CNTs was 100:10:1).

    Techniques: Irradiation

    Top Panel: SDS-PAGE analysis of time course digestion of Vip3Ab1 and Vip3Bc1 with H. zea and P. includens gut enzymes. Vip3Ab1 and Vip3Bc1 proteins (150 µg/mL) were incubated with gut fluids from H. zea (left) and P. includens (right) at 30 °C for various time intervals at pH 10.0. Bottom Panel: SDS-PAGE analysis of overnight digestion of Vip3 chimeras with H. zea gut enzymes. Vip3_AB and Vip3_BA proteins (110 µg/ml) were incubated with H. zea gut fluids for 16 hours at 30 °C in a total volume of 100 µL at pH 10.0. All reactions were stopped with protease inhibitors and 30 µL of the reaction loaded as described in Materials and Methods. Equivalent lanes were loaded and blotted onto a PVDF membrane for N-terminal sequencing.

    Journal: Scientific Reports

    Article Title: Functional characterization of Vip3Ab1 and Vip3Bc1: Two novel insecticidal proteins with differential activity against lepidopteran pests

    doi: 10.1038/s41598-017-11702-2

    Figure Lengend Snippet: Top Panel: SDS-PAGE analysis of time course digestion of Vip3Ab1 and Vip3Bc1 with H. zea and P. includens gut enzymes. Vip3Ab1 and Vip3Bc1 proteins (150 µg/mL) were incubated with gut fluids from H. zea (left) and P. includens (right) at 30 °C for various time intervals at pH 10.0. Bottom Panel: SDS-PAGE analysis of overnight digestion of Vip3 chimeras with H. zea gut enzymes. Vip3_AB and Vip3_BA proteins (110 µg/ml) were incubated with H. zea gut fluids for 16 hours at 30 °C in a total volume of 100 µL at pH 10.0. All reactions were stopped with protease inhibitors and 30 µL of the reaction loaded as described in Materials and Methods. Equivalent lanes were loaded and blotted onto a PVDF membrane for N-terminal sequencing.

    Article Snippet: Samples were prepared for N-terminal sequencing by blotting an SDS-PAGE gel onto a PVDF pre-cut blotting membrane (Thermo Scientific, Waltham, MA) via wet tank transfer in 10 mM CAPS (pH 11) with 10% methanol.

    Techniques: SDS Page, Incubation, Sequencing