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    Name:
    QIAquick Gel Extraction Kit
    Description:
    For gel extraction cleanup of up to 10 μg DNA 70 bp to 10 kb from enzymatic reactions Kit contents Qiagen QIAquick Gel Extraction Kit 50 rxns 30 to 50L Elution Volume 10g Binding Capacity DNA Sample Tube Format Silica Technology Manual Processing 70 bp to 10 kb Fragment Fast and Convenient Procedure For Gel Extraction Cleanup of up to 10μg DNA 70 bp to 10 kb from Enzymatic Reactions Includes 50 QIAquick Spin Columns Buffers Collection Tubes 2mL Benefits Up to 95 recovery of ready to use DNA Fast and convenient procedure Cleanup of DNA up to 10 kb in three easy steps Gel loading dye for convenient sample analysis
    Catalog Number:
    28704
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    QIAquick Gel Extraction Kit
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    Qiagen hepcs
    QIAquick Gel Extraction Kit
    For gel extraction cleanup of up to 10 μg DNA 70 bp to 10 kb from enzymatic reactions Kit contents Qiagen QIAquick Gel Extraction Kit 50 rxns 30 to 50L Elution Volume 10g Binding Capacity DNA Sample Tube Format Silica Technology Manual Processing 70 bp to 10 kb Fragment Fast and Convenient Procedure For Gel Extraction Cleanup of up to 10μg DNA 70 bp to 10 kb from Enzymatic Reactions Includes 50 QIAquick Spin Columns Buffers Collection Tubes 2mL Benefits Up to 95 recovery of ready to use DNA Fast and convenient procedure Cleanup of DNA up to 10 kb in three easy steps Gel loading dye for convenient sample analysis
    https://www.bioz.com/result/hepcs/product/Qiagen
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hepcs - by Bioz Stars, 2021-03
    99/100 stars

    Images

    1) Product Images from "Loss of Apelin Exacerbates Myocardial Infarction Adverse Remodeling and Ischemia-reperfusion Injury: Therapeutic Potential of Synthetic Apelin Analogues"

    Article Title: Loss of Apelin Exacerbates Myocardial Infarction Adverse Remodeling and Ischemia-reperfusion Injury: Therapeutic Potential of Synthetic Apelin Analogues

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    doi: 10.1161/JAHA.113.000249

    Loss of apelin impairs the in vitro angiogenic response in murine aorta and human endothelial progenitor cells while apelin analogue stimulates angiogenesis. (A) Representative aortic ring cultures from APLN +/y and APLN ‐/y mice showing a marked decrease in sprout length and density which was rescued by supplementation with apelin analogue II (100 ng/mL) and rhVEGF (20 ng/mL). (B) Characterization of the isolated human endothelial progenitor cells (hEPCs) using flow cytometry of the endothelial‐specific marker, VE‐cadherin while Western blot analysis of von Willebrand factor (vWF) and eNOS showed comparable expression with a positive control, human umbilical vein endothelial cells (HUVEC). Transfection of human endothelial progenitor cells with a non‐silencing control siRNA (siNS) or siRNA against apelin (siApelin) resulting in 80% decrease in apelin (C) and DLL4 (D) expression. (E) Representative beads and quantification of sprout length based on tertiles established from mock‐transfected hEPC (Mock) and sprout density showing a drastic reduction in endothelial sprout density and sprout lengthening with knockdown of apelin. Apelin analogue II (100 ng/mL) stimulates endothelial lengthening (E) and increased phospho‐Akt (serine‐473) in hEPCs (F). rhVEGF indicates recombinant human vascular endothelial growth factor; APLN, Apelin; NS, nonsense; p, phospho; t, total; R.R., relative ratio; eNOS, endothelial nitric oxide synthase. Values are mean±SEM; n=5 for each group. * P
    Figure Legend Snippet: Loss of apelin impairs the in vitro angiogenic response in murine aorta and human endothelial progenitor cells while apelin analogue stimulates angiogenesis. (A) Representative aortic ring cultures from APLN +/y and APLN ‐/y mice showing a marked decrease in sprout length and density which was rescued by supplementation with apelin analogue II (100 ng/mL) and rhVEGF (20 ng/mL). (B) Characterization of the isolated human endothelial progenitor cells (hEPCs) using flow cytometry of the endothelial‐specific marker, VE‐cadherin while Western blot analysis of von Willebrand factor (vWF) and eNOS showed comparable expression with a positive control, human umbilical vein endothelial cells (HUVEC). Transfection of human endothelial progenitor cells with a non‐silencing control siRNA (siNS) or siRNA against apelin (siApelin) resulting in 80% decrease in apelin (C) and DLL4 (D) expression. (E) Representative beads and quantification of sprout length based on tertiles established from mock‐transfected hEPC (Mock) and sprout density showing a drastic reduction in endothelial sprout density and sprout lengthening with knockdown of apelin. Apelin analogue II (100 ng/mL) stimulates endothelial lengthening (E) and increased phospho‐Akt (serine‐473) in hEPCs (F). rhVEGF indicates recombinant human vascular endothelial growth factor; APLN, Apelin; NS, nonsense; p, phospho; t, total; R.R., relative ratio; eNOS, endothelial nitric oxide synthase. Values are mean±SEM; n=5 for each group. * P

    Techniques Used: In Vitro, Mouse Assay, Isolation, Flow Cytometry, Cytometry, Marker, Western Blot, Expressing, Positive Control, Transfection, Recombinant

    2) Product Images from "Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation"

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation

    Journal: Peptides

    doi: 10.1016/j.peptides.2008.08.006

    Cadherin peptides enhance Cry1Ac (black bars) and Cry1Ab (empty bars) toxins when fed to M. sexta larvae. LC10 concentration of Cry1Ac or Cry1Ab toxins (0.05 ng/cm 2 of diet) were mixed with 5ng/cm 2 of different cadherin peptides corresponding to CR7-12, CR12, CR11, CR7, CR7-12-I1422R and CR12-I1422R. Mortality % of 48 larvae after five days in diet. Representative results of two experiments are shown.
    Figure Legend Snippet: Cadherin peptides enhance Cry1Ac (black bars) and Cry1Ab (empty bars) toxins when fed to M. sexta larvae. LC10 concentration of Cry1Ac or Cry1Ab toxins (0.05 ng/cm 2 of diet) were mixed with 5ng/cm 2 of different cadherin peptides corresponding to CR7-12, CR12, CR11, CR7, CR7-12-I1422R and CR12-I1422R. Mortality % of 48 larvae after five days in diet. Representative results of two experiments are shown.

    Techniques Used: Concentration Assay

    Oligomer formation of Cry1Ab protoxin in the presence of different cadherin fragments. A. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR7, CR11 or CR12 and oligomer formation was revealed after SDS-PAGE electrophoresis and Western blot using anti-Cry1Ab antibody. NR is without any CR peptide. B. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR12 and CR12-I1422R and oligomer were revealed as described above.
    Figure Legend Snippet: Oligomer formation of Cry1Ab protoxin in the presence of different cadherin fragments. A. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR7, CR11 or CR12 and oligomer formation was revealed after SDS-PAGE electrophoresis and Western blot using anti-Cry1Ab antibody. NR is without any CR peptide. B. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR12 and CR12-I1422R and oligomer were revealed as described above.

    Techniques Used: Incubation, SDS Page, Electrophoresis, Western Blot

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation
    Article Snippet: The PCR reactions were performed with Vent-Polymerase (New England BioLabs, Beverly, MA). .. The PCR products (342 bp for CR7 and 429 pb for CR11) were purified with QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), digested with EcoRI and HindIII (New England BioLabs, Beverly, MA) and ligated into the previously digested vector pET22b (Novagen, EMD Biosciences, Inc.). .. All cadherin fragments including the CR7-12 and CR12 fragments previously cloned [ , ] were purified to homogeneity from BL21 E. coli cultures grown at 37 °C in 2X TY (Tryptone 1.6 % w/v, Yeast extract 1% w/v and NaCl 0.5 % w/v, supplemented with 100 μg/ml ampicillin and 0.1% glucose) until they reached an OD of 0.7 at 600 nm.

    Article Title: Experimental infection of horses with Rickettsia rickettsii
    Article Snippet: Positive and negative control sera corresponding to each animal species were added to each IFA slide. .. PCR DNA extraction from horse blood and guinea pig organs was performed with the commercial kit DNeasy Blood & Tissue Kit (QIAGEN, Hilden, Germany), according to the manufacturer’s instructions. ..

    Article Title: In vivo delivery of transcription factors with multifunctional oligonucleotides
    Article Snippet: For all in vitro studies, 0.6 μM of Nrf2 or 0.6 μM of Nrf2 complexed with 0.6 μM of DARTs were used. .. Reverse transcription PCR (RT-PCR) analysis of HepG2 cells treated with Nrf2 delivered by DARTs HepG2 cells were treated with either free Nrf2 or DART-Nrf2 in culture media for 24 h and the RNA was extracted from the cells using the RNeasy® Mini Kit with additional DNase I digestion (Qiagen, Hilden, Germany), according to the manufacturer’s protocol. .. RNA purity and concentration were determined by measuring the optical density using an ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE, USA).

    Purification:

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation
    Article Snippet: The PCR reactions were performed with Vent-Polymerase (New England BioLabs, Beverly, MA). .. The PCR products (342 bp for CR7 and 429 pb for CR11) were purified with QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), digested with EcoRI and HindIII (New England BioLabs, Beverly, MA) and ligated into the previously digested vector pET22b (Novagen, EMD Biosciences, Inc.). .. All cadherin fragments including the CR7-12 and CR12 fragments previously cloned [ , ] were purified to homogeneity from BL21 E. coli cultures grown at 37 °C in 2X TY (Tryptone 1.6 % w/v, Yeast extract 1% w/v and NaCl 0.5 % w/v, supplemented with 100 μg/ml ampicillin and 0.1% glucose) until they reached an OD of 0.7 at 600 nm.

    Plasmid Preparation:

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation
    Article Snippet: The PCR reactions were performed with Vent-Polymerase (New England BioLabs, Beverly, MA). .. The PCR products (342 bp for CR7 and 429 pb for CR11) were purified with QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), digested with EcoRI and HindIII (New England BioLabs, Beverly, MA) and ligated into the previously digested vector pET22b (Novagen, EMD Biosciences, Inc.). .. All cadherin fragments including the CR7-12 and CR12 fragments previously cloned [ , ] were purified to homogeneity from BL21 E. coli cultures grown at 37 °C in 2X TY (Tryptone 1.6 % w/v, Yeast extract 1% w/v and NaCl 0.5 % w/v, supplemented with 100 μg/ml ampicillin and 0.1% glucose) until they reached an OD of 0.7 at 600 nm.

    Transfection:

    Article Title: Loss of Apelin Exacerbates Myocardial Infarction Adverse Remodeling and Ischemia-reperfusion Injury: Therapeutic Potential of Synthetic Apelin Analogues
    Article Snippet: The late outgrowth endothelial colony‐forming cells (ECFC) clones were expanded in EBM‐2 medium with 10% FBS, and routinely monitored for expression of endothelial marker gene products using quantitative RT‐PCR, flow cytometry, or Western blot. .. Where indicated, hEPCs were transfected with 50 mmol/L non‐silencing or siRNA directed to APLN (Qiagen) and in vitro angiogenesis of hEPCs was evaluated as described previously. .. – EPCs transfected with si Nonsense (siNS) or siAPLN were loaded onto Cytodex (Sigma) beads (~400 cells/bead) and cultured for 2 hours, then the beads were suspended in fibrinogen/fibronectin solution (2 mg/mL) containing aprotinin (0.15 U/mL) and 0.625 U/mL thrombin was added.

    In Vitro:

    Article Title: Loss of Apelin Exacerbates Myocardial Infarction Adverse Remodeling and Ischemia-reperfusion Injury: Therapeutic Potential of Synthetic Apelin Analogues
    Article Snippet: The late outgrowth endothelial colony‐forming cells (ECFC) clones were expanded in EBM‐2 medium with 10% FBS, and routinely monitored for expression of endothelial marker gene products using quantitative RT‐PCR, flow cytometry, or Western blot. .. Where indicated, hEPCs were transfected with 50 mmol/L non‐silencing or siRNA directed to APLN (Qiagen) and in vitro angiogenesis of hEPCs was evaluated as described previously. .. – EPCs transfected with si Nonsense (siNS) or siAPLN were loaded onto Cytodex (Sigma) beads (~400 cells/bead) and cultured for 2 hours, then the beads were suspended in fibrinogen/fibronectin solution (2 mg/mL) containing aprotinin (0.15 U/mL) and 0.625 U/mL thrombin was added.

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.
    Article Snippet: Cleavage assay was performed as we have previously described using recombinant His tagged VWF-A2 protein with modifications [ ]. .. Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate. .. Microtiter wells were incubated with diluted plasma for 16 hours.

    DNA Extraction:

    Article Title: Experimental infection of horses with Rickettsia rickettsii
    Article Snippet: Positive and negative control sera corresponding to each animal species were added to each IFA slide. .. PCR DNA extraction from horse blood and guinea pig organs was performed with the commercial kit DNeasy Blood & Tissue Kit (QIAGEN, Hilden, Germany), according to the manufacturer’s instructions. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: In vivo delivery of transcription factors with multifunctional oligonucleotides
    Article Snippet: For all in vitro studies, 0.6 μM of Nrf2 or 0.6 μM of Nrf2 complexed with 0.6 μM of DARTs were used. .. Reverse transcription PCR (RT-PCR) analysis of HepG2 cells treated with Nrf2 delivered by DARTs HepG2 cells were treated with either free Nrf2 or DART-Nrf2 in culture media for 24 h and the RNA was extracted from the cells using the RNeasy® Mini Kit with additional DNase I digestion (Qiagen, Hilden, Germany), according to the manufacturer’s protocol. .. RNA purity and concentration were determined by measuring the optical density using an ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE, USA).

    Isolation:

    Article Title: T cell-intrinsic IL-1R signaling licenses effector cytokine production by memory CD4 T cells
    Article Snippet: At given time points after addition of actinomycin D cells were washed with cold PBS and lysed with TriZol (Life Technologies) and frozen until further processing. .. RNA was isolated using Qiagen RNA extraction kit using manufacturer’s protocol. cDNA was synthesized using Random primers (Invitrogen) and MMLV reverse transcriptase (Invitrogen). .. The QuantStudio 7 Flex Real-Time PCR System (ThermoFisher Scientific) was used to measure SGBR green (ThermoFisher Scientific) incorporation.

    RNA Extraction:

    Article Title: T cell-intrinsic IL-1R signaling licenses effector cytokine production by memory CD4 T cells
    Article Snippet: At given time points after addition of actinomycin D cells were washed with cold PBS and lysed with TriZol (Life Technologies) and frozen until further processing. .. RNA was isolated using Qiagen RNA extraction kit using manufacturer’s protocol. cDNA was synthesized using Random primers (Invitrogen) and MMLV reverse transcriptase (Invitrogen). .. The QuantStudio 7 Flex Real-Time PCR System (ThermoFisher Scientific) was used to measure SGBR green (ThermoFisher Scientific) incorporation.

    Synthesized:

    Article Title: T cell-intrinsic IL-1R signaling licenses effector cytokine production by memory CD4 T cells
    Article Snippet: At given time points after addition of actinomycin D cells were washed with cold PBS and lysed with TriZol (Life Technologies) and frozen until further processing. .. RNA was isolated using Qiagen RNA extraction kit using manufacturer’s protocol. cDNA was synthesized using Random primers (Invitrogen) and MMLV reverse transcriptase (Invitrogen). .. The QuantStudio 7 Flex Real-Time PCR System (ThermoFisher Scientific) was used to measure SGBR green (ThermoFisher Scientific) incorporation.

    Kinase Assay:

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.
    Article Snippet: Cleavage assay was performed as we have previously described using recombinant His tagged VWF-A2 protein with modifications [ ]. .. Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate. .. Microtiter wells were incubated with diluted plasma for 16 hours.

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    Qiagen in vitro fam20c kinase assay
    A1A2A3 protein phosphorylated at S1613 enhances platelet-VWF interaction. ( A ) Coomassie staining of the purified A1A2A3 wild type (WT) and A1A2A3 S1517A mutant. ( B ) Whole blood was perfused over immobilized A1A2A3 (WT), A1A2A3 (S1517A) mutant that was previously incubated with <t>FAM20c</t> in the presence of ATP (S1613 phosphorylated) and absence of ATP (S1613 not phosphorylated) at a shear rate of 1500 S −1 and adhesion visualized in a Bioflux system. ( C ) Quantification of platelet coverage and stable microthrombi. Data are obtained from six independent blood donors.
    In Vitro Fam20c Kinase Assay, supplied by Qiagen, 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/in vitro fam20c kinase assay/product/Qiagen
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    in vitro fam20c kinase assay - by Bioz Stars, 2021-03
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    A1A2A3 protein phosphorylated at S1613 enhances platelet-VWF interaction. ( A ) Coomassie staining of the purified A1A2A3 wild type (WT) and A1A2A3 S1517A mutant. ( B ) Whole blood was perfused over immobilized A1A2A3 (WT), A1A2A3 (S1517A) mutant that was previously incubated with FAM20c in the presence of ATP (S1613 phosphorylated) and absence of ATP (S1613 not phosphorylated) at a shear rate of 1500 S −1 and adhesion visualized in a Bioflux system. ( C ) Quantification of platelet coverage and stable microthrombi. Data are obtained from six independent blood donors.

    Journal: Journal of thrombosis and haemostasis : JTH

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.

    doi: 10.1111/jth.14426

    Figure Lengend Snippet: A1A2A3 protein phosphorylated at S1613 enhances platelet-VWF interaction. ( A ) Coomassie staining of the purified A1A2A3 wild type (WT) and A1A2A3 S1517A mutant. ( B ) Whole blood was perfused over immobilized A1A2A3 (WT), A1A2A3 (S1517A) mutant that was previously incubated with FAM20c in the presence of ATP (S1613 phosphorylated) and absence of ATP (S1613 not phosphorylated) at a shear rate of 1500 S −1 and adhesion visualized in a Bioflux system. ( C ) Quantification of platelet coverage and stable microthrombi. Data are obtained from six independent blood donors.

    Article Snippet: Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate.

    Techniques: Staining, Purification, Mutagenesis, Incubation

    Identification of S1517 and S1613 in the VWF A2 protein as FAM20c phosphorylation sites. Mass spectrometric analysis of the A2 protein subjected to FAM20c in vitro kinase assays. ( A) LC/MS e low energy (MS 1 ) precursor ion mass spectra showing A2 domain protein incubated with FAM20C kinase in the absence and presence of ATP. Lock mass corrected and charge-reduced, mass-centroid base peak spectra are shown where x-axis represents M+H + (Da), and y-axis is relative ion intensities; numbers shown on top of spectrum peaks represent LC retention times. Peaks circled in the +ATP spectrum show the presence of phosphorylated peptides not found in the -ATP spectrum, indicating FAM20c-mediated specific phosphorylation. Phosphopeptides in circle i and circle ii correspond to phosphopeptide isoforms which include FAM20c-mediated phosphorylation on Ser-1517 and those in circle iii and iv to phosphopeptide isoforms which include FAM20c-mediated phosphorylation on Ser-1613. ( B) LC/MS E base peak high energy (MS 2 ) product ion mass spectra showing all b- and y-ions observed in the MS2 spectrum for two of the phosphopeptide isoforms circled in ii and iv respectively. These MS2 spectra show unequivocally (as summarized in the peptide sequence display shown at the top of each spectra) the site of FAM20c- mediated phosphorylation on the A2 domain protein as being localized to Ser-1517 and Ser-1613. pS refers to site of phosphorylation, and oxM refers to an oxidized methionine residue. (C) Incorporation of 32 P from γ− 32 P ATP to wild type (WT) VWF A2 and various A2 mutants by FAM20c and FAM20c kinase dead mutant. Samples were separated by SDS-PAGE and subjected to autoradiography (upper panel) or visualized by Coomassie blue staining (lower panel). Blots are representative of two independent experiments.

    Journal: Journal of thrombosis and haemostasis : JTH

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.

    doi: 10.1111/jth.14426

    Figure Lengend Snippet: Identification of S1517 and S1613 in the VWF A2 protein as FAM20c phosphorylation sites. Mass spectrometric analysis of the A2 protein subjected to FAM20c in vitro kinase assays. ( A) LC/MS e low energy (MS 1 ) precursor ion mass spectra showing A2 domain protein incubated with FAM20C kinase in the absence and presence of ATP. Lock mass corrected and charge-reduced, mass-centroid base peak spectra are shown where x-axis represents M+H + (Da), and y-axis is relative ion intensities; numbers shown on top of spectrum peaks represent LC retention times. Peaks circled in the +ATP spectrum show the presence of phosphorylated peptides not found in the -ATP spectrum, indicating FAM20c-mediated specific phosphorylation. Phosphopeptides in circle i and circle ii correspond to phosphopeptide isoforms which include FAM20c-mediated phosphorylation on Ser-1517 and those in circle iii and iv to phosphopeptide isoforms which include FAM20c-mediated phosphorylation on Ser-1613. ( B) LC/MS E base peak high energy (MS 2 ) product ion mass spectra showing all b- and y-ions observed in the MS2 spectrum for two of the phosphopeptide isoforms circled in ii and iv respectively. These MS2 spectra show unequivocally (as summarized in the peptide sequence display shown at the top of each spectra) the site of FAM20c- mediated phosphorylation on the A2 domain protein as being localized to Ser-1517 and Ser-1613. pS refers to site of phosphorylation, and oxM refers to an oxidized methionine residue. (C) Incorporation of 32 P from γ− 32 P ATP to wild type (WT) VWF A2 and various A2 mutants by FAM20c and FAM20c kinase dead mutant. Samples were separated by SDS-PAGE and subjected to autoradiography (upper panel) or visualized by Coomassie blue staining (lower panel). Blots are representative of two independent experiments.

    Article Snippet: Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate.

    Techniques: In Vitro, Liquid Chromatography with Mass Spectroscopy, Incubation, Sequencing, Mutagenesis, SDS Page, Autoradiography, Staining

    In vitro phosphorylation of VWF A domains by FAM20c. Incorporation of 32 P from γ− 32 P ATP to VWF A1A2A3 protein ( A ) and the individual A domains ( B ) by FAM20c but not by the FAM20c kinase dead mutant (D478A). Samples from kinase assays were separated by SDS-PAGE and subjected to autoradiography (upper panel) or visualized by Coomasssie blue staining (lower panel). Degradation product of A1 protein in Figure 1B is shown as **. Phosphorylation of A1A2A3 was blocked by treatment with λ phosphatase. Autophosphorylation of FAM20c in this assay is labelled as FAM20c. Blots are representative of three independent experiments.

    Journal: Journal of thrombosis and haemostasis : JTH

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.

    doi: 10.1111/jth.14426

    Figure Lengend Snippet: In vitro phosphorylation of VWF A domains by FAM20c. Incorporation of 32 P from γ− 32 P ATP to VWF A1A2A3 protein ( A ) and the individual A domains ( B ) by FAM20c but not by the FAM20c kinase dead mutant (D478A). Samples from kinase assays were separated by SDS-PAGE and subjected to autoradiography (upper panel) or visualized by Coomasssie blue staining (lower panel). Degradation product of A1 protein in Figure 1B is shown as **. Phosphorylation of A1A2A3 was blocked by treatment with λ phosphatase. Autophosphorylation of FAM20c in this assay is labelled as FAM20c. Blots are representative of three independent experiments.

    Article Snippet: Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate.

    Techniques: In Vitro, Mutagenesis, SDS Page, Autoradiography, Staining

    Increased platelet adhesion to A1A2A3 protein from FAM20c transfected cells (A) Immunoblotting of A1A2A3 protein purified from HEK 293 cells that are transfected with A1A2A3 and dead FAM20c kinase mutant D478A (D478A) or A1A2A3 and FAM20c (FAM20c) with anti-VWF antibody. (B) Whole blood was perfused over A1A2A3 protein obtained from cells with D478A or FAM20c at a shear rate of 1500 S −1 and adhesion visualized in a Bioflux system. ( C ) Quantification of platelet coverage is depicted in a logarithmic scale. Data are obtained from three independent blood donors.

    Journal: Journal of thrombosis and haemostasis : JTH

    Article Title: In vitro phosphorylation of von Willebrand factor by FAM20c enhances its ability to support platelet adhesion.

    doi: 10.1111/jth.14426

    Figure Lengend Snippet: Increased platelet adhesion to A1A2A3 protein from FAM20c transfected cells (A) Immunoblotting of A1A2A3 protein purified from HEK 293 cells that are transfected with A1A2A3 and dead FAM20c kinase mutant D478A (D478A) or A1A2A3 and FAM20c (FAM20c) with anti-VWF antibody. (B) Whole blood was perfused over A1A2A3 protein obtained from cells with D478A or FAM20c at a shear rate of 1500 S −1 and adhesion visualized in a Bioflux system. ( C ) Quantification of platelet coverage is depicted in a logarithmic scale. Data are obtained from three independent blood donors.

    Article Snippet: Briefly, A2 protein subjected to in vitro FAM20c kinase assay in the presence or absence of ATP was absorbed to Ni-NTA HisSorb strips (Qiagen) as substrate.

    Techniques: Transfection, Purification, Mutagenesis

    Loss of apelin impairs the in vitro angiogenic response in murine aorta and human endothelial progenitor cells while apelin analogue stimulates angiogenesis. (A) Representative aortic ring cultures from APLN +/y and APLN ‐/y mice showing a marked decrease in sprout length and density which was rescued by supplementation with apelin analogue II (100 ng/mL) and rhVEGF (20 ng/mL). (B) Characterization of the isolated human endothelial progenitor cells (hEPCs) using flow cytometry of the endothelial‐specific marker, VE‐cadherin while Western blot analysis of von Willebrand factor (vWF) and eNOS showed comparable expression with a positive control, human umbilical vein endothelial cells (HUVEC). Transfection of human endothelial progenitor cells with a non‐silencing control siRNA (siNS) or siRNA against apelin (siApelin) resulting in 80% decrease in apelin (C) and DLL4 (D) expression. (E) Representative beads and quantification of sprout length based on tertiles established from mock‐transfected hEPC (Mock) and sprout density showing a drastic reduction in endothelial sprout density and sprout lengthening with knockdown of apelin. Apelin analogue II (100 ng/mL) stimulates endothelial lengthening (E) and increased phospho‐Akt (serine‐473) in hEPCs (F). rhVEGF indicates recombinant human vascular endothelial growth factor; APLN, Apelin; NS, nonsense; p, phospho; t, total; R.R., relative ratio; eNOS, endothelial nitric oxide synthase. Values are mean±SEM; n=5 for each group. * P

    Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

    Article Title: Loss of Apelin Exacerbates Myocardial Infarction Adverse Remodeling and Ischemia-reperfusion Injury: Therapeutic Potential of Synthetic Apelin Analogues

    doi: 10.1161/JAHA.113.000249

    Figure Lengend Snippet: Loss of apelin impairs the in vitro angiogenic response in murine aorta and human endothelial progenitor cells while apelin analogue stimulates angiogenesis. (A) Representative aortic ring cultures from APLN +/y and APLN ‐/y mice showing a marked decrease in sprout length and density which was rescued by supplementation with apelin analogue II (100 ng/mL) and rhVEGF (20 ng/mL). (B) Characterization of the isolated human endothelial progenitor cells (hEPCs) using flow cytometry of the endothelial‐specific marker, VE‐cadherin while Western blot analysis of von Willebrand factor (vWF) and eNOS showed comparable expression with a positive control, human umbilical vein endothelial cells (HUVEC). Transfection of human endothelial progenitor cells with a non‐silencing control siRNA (siNS) or siRNA against apelin (siApelin) resulting in 80% decrease in apelin (C) and DLL4 (D) expression. (E) Representative beads and quantification of sprout length based on tertiles established from mock‐transfected hEPC (Mock) and sprout density showing a drastic reduction in endothelial sprout density and sprout lengthening with knockdown of apelin. Apelin analogue II (100 ng/mL) stimulates endothelial lengthening (E) and increased phospho‐Akt (serine‐473) in hEPCs (F). rhVEGF indicates recombinant human vascular endothelial growth factor; APLN, Apelin; NS, nonsense; p, phospho; t, total; R.R., relative ratio; eNOS, endothelial nitric oxide synthase. Values are mean±SEM; n=5 for each group. * P

    Article Snippet: Where indicated, hEPCs were transfected with 50 mmol/L non‐silencing or siRNA directed to APLN (Qiagen) and in vitro angiogenesis of hEPCs was evaluated as described previously.

    Techniques: In Vitro, Mouse Assay, Isolation, Flow Cytometry, Cytometry, Marker, Western Blot, Expressing, Positive Control, Transfection, Recombinant

    Cadherin peptides enhance Cry1Ac (black bars) and Cry1Ab (empty bars) toxins when fed to M. sexta larvae. LC10 concentration of Cry1Ac or Cry1Ab toxins (0.05 ng/cm 2 of diet) were mixed with 5ng/cm 2 of different cadherin peptides corresponding to CR7-12, CR12, CR11, CR7, CR7-12-I1422R and CR12-I1422R. Mortality % of 48 larvae after five days in diet. Representative results of two experiments are shown.

    Journal: Peptides

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation

    doi: 10.1016/j.peptides.2008.08.006

    Figure Lengend Snippet: Cadherin peptides enhance Cry1Ac (black bars) and Cry1Ab (empty bars) toxins when fed to M. sexta larvae. LC10 concentration of Cry1Ac or Cry1Ab toxins (0.05 ng/cm 2 of diet) were mixed with 5ng/cm 2 of different cadherin peptides corresponding to CR7-12, CR12, CR11, CR7, CR7-12-I1422R and CR12-I1422R. Mortality % of 48 larvae after five days in diet. Representative results of two experiments are shown.

    Article Snippet: The PCR products (342 bp for CR7 and 429 pb for CR11) were purified with QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), digested with EcoRI and HindIII (New England BioLabs, Beverly, MA) and ligated into the previously digested vector pET22b (Novagen, EMD Biosciences, Inc.).

    Techniques: Concentration Assay

    Oligomer formation of Cry1Ab protoxin in the presence of different cadherin fragments. A. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR7, CR11 or CR12 and oligomer formation was revealed after SDS-PAGE electrophoresis and Western blot using anti-Cry1Ab antibody. NR is without any CR peptide. B. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR12 and CR12-I1422R and oligomer were revealed as described above.

    Journal: Peptides

    Article Title: Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxin-binding cadherin correlates with oligomer formation

    doi: 10.1016/j.peptides.2008.08.006

    Figure Lengend Snippet: Oligomer formation of Cry1Ab protoxin in the presence of different cadherin fragments. A. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR7, CR11 or CR12 and oligomer formation was revealed after SDS-PAGE electrophoresis and Western blot using anti-Cry1Ab antibody. NR is without any CR peptide. B. Cry1Ab protoxin was incubated with 5% midgut juice in the presence of CR12 and CR12-I1422R and oligomer were revealed as described above.

    Article Snippet: The PCR products (342 bp for CR7 and 429 pb for CR11) were purified with QIAquick PCR Purification Kit (QIAGEN, Valencia, CA), digested with EcoRI and HindIII (New England BioLabs, Beverly, MA) and ligated into the previously digested vector pET22b (Novagen, EMD Biosciences, Inc.).

    Techniques: Incubation, SDS Page, Electrophoresis, Western Blot

    DARTs are able to deliver Nrf2 to hepatocytes, up-regulate Nrf2 downstream genes, and can protect hepatocytes against reactive oxygen species (ROS) a , Nrf2 delivered by DARTs enhances the expression of HO1, NQO1, and GCLC. RT-PCR of HepG2 cells treated with DART-Nrf2 complexes up-regulate HO1, NQO1, and GCLC. Free Nrf2 had no effect on HO1, NQO1, and GCLC gene expression. b , Nrf2 delivered by DARTs reduces ROS levels in HepG2 cells stressed with hydrogen peroxide. ROS levels in hydrogen peroxide stressed cells were not significantly reduced by free Nrf2, whereas DART-Nrf2 decreased ROS production down to levels comparable to control cells, mean ± S.E, n=9. **, p

    Journal: Nature materials

    Article Title: In vivo delivery of transcription factors with multifunctional oligonucleotides

    doi: 10.1038/nmat4269

    Figure Lengend Snippet: DARTs are able to deliver Nrf2 to hepatocytes, up-regulate Nrf2 downstream genes, and can protect hepatocytes against reactive oxygen species (ROS) a , Nrf2 delivered by DARTs enhances the expression of HO1, NQO1, and GCLC. RT-PCR of HepG2 cells treated with DART-Nrf2 complexes up-regulate HO1, NQO1, and GCLC. Free Nrf2 had no effect on HO1, NQO1, and GCLC gene expression. b , Nrf2 delivered by DARTs reduces ROS levels in HepG2 cells stressed with hydrogen peroxide. ROS levels in hydrogen peroxide stressed cells were not significantly reduced by free Nrf2, whereas DART-Nrf2 decreased ROS production down to levels comparable to control cells, mean ± S.E, n=9. **, p

    Article Snippet: Reverse transcription PCR (RT-PCR) analysis of HepG2 cells treated with Nrf2 delivered by DARTs HepG2 cells were treated with either free Nrf2 or DART-Nrf2 in culture media for 24 h and the RNA was extracted from the cells using the RNeasy® Mini Kit with additional DNase I digestion (Qiagen, Hilden, Germany), according to the manufacturer’s protocol.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction