ribonuclease a rnasea  (Millipore)


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    Name:
    Ribonuclease A from bovine pancreas
    Description:
    Ribonucleases do not hydrolyze DNA because the DNA lacks 2 OH groups essential for the formation of cyclic intermediates RNase can hydrolyze RNA from protein samples Pancreatic RNase A specifically cleaves at the 3 side of pyrimidine uracil or cytosine phosphate bonds
    Catalog Number:
    r5000
    Price:
    None
    Applications:
    Ribonuclease A is used to remove RNA from DNA plasmid preparations and protein samples. Ribonuclease A is used for RNase protection assays, to remove unspecifically bound RNA, analysis of RNA sequences, to hydrolyze RNA contained in protein samples, and the purification of DNA. Ribonuclease A from bovine pancreas has been used in a study to assess hybridase activity of human ribonuclease-1. Ribonuclease A from bovine pancreas has also been used in a study to investigate particle-based and monolithic columns for cation exchange protein displacement chromatography.
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    Structured Review

    Millipore ribonuclease a rnasea
    Ribonuclease A from bovine pancreas
    Ribonucleases do not hydrolyze DNA because the DNA lacks 2 OH groups essential for the formation of cyclic intermediates RNase can hydrolyze RNA from protein samples Pancreatic RNase A specifically cleaves at the 3 side of pyrimidine uracil or cytosine phosphate bonds
    https://www.bioz.com/result/ribonuclease a rnasea/product/Millipore
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    ribonuclease a rnasea - by Bioz Stars, 2020-09
    99/100 stars

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    Avidin-Biotin Assay:

    Article Title: Net charge of antibody complementarity-determining regions is a key predictor of specificity
    Article Snippet: .. The panel of proteins included ovalbumin (pI = 4.6; S25132, Thermo Fisher Scientific), bovine serum albumin (BSA; pI = 4.7; BP9706 Thermo Fisher Scientific), insulin (pI = 5.4; I9278, Sigma), keyhole limpet hemocyanin (KLH; pI = 4.6; H8283, Sigma), ribonuclease A (pI = 9.3; R4875, Sigma), avidin (pI = 10; A9275, Sigma) and lysozyme (pI = 11, L6876, Sigma). ..

    Adsorption:

    Article Title: Adsorption-Induced Changes in Ribonuclease A Structure and Enzymatic Activity on Solid Surfaces
    Article Snippet: .. II.b Protein Adsorption and Equilibration The adsorption of RNAse A (Sigma R6513) on the material surfaces was carried out using previously described methods (see S.1.a in the ). .. Briefly, 10 mM potassium phosphate buffer solution (PPB; pH 7.4) was prepared by mixing appropriate amounts of 1 M monobasic potassium phosphate (Sigma, P8708) or 1 M dibasic potassium phosphate (Sigma, P8508), following which the buffer concentration was verified by titrating against 0.065 M potassium hydrogen phthalate.

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  • 85
    Millipore myc rac1
    Effect of AA and SDS on the interaction of the p67 phox ·Rac-GTP complex with Nox2. A , SDS-PAGE analysis of purified proteins used in this study. GST alone, GST-Nox2-C, the wild-type p67 phox <t>(1–212)-Rac1</t> (Q61L) chimera, and a mutant p67
    Myc Rac1, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/myc rac1/product/Millipore
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    myc rac1 - by Bioz Stars, 2020-09
    85/100 stars
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    90
    Millipore gtp rac1 pull down assay
    CRKL mediates ALK signaling and regulates cytoskeleton, cell migration and survival A. CRKL siRNA knockdown in H2228 and H3122. Protein lysates were extracted from cells treated with four individual CRLK siRNAs (20 nM) or their smartpool for 72 h, and subjected to Western blot analyses. The graph shows the quantification of CRKL proteins. B. Inhibition of Ras GTPase activity by CRKL siRNA knockdown. Upper left panel: Lysates of untreated H3122 cells were incubated with buffer (Ctrl), non-hydrolyzable analog of <t>GTP</t> (GTPγS as a positive control) or GDP (as a negative control). Upper right panel: Lysates of H3122 cells treated with or without CRKL siRNA were incubated with GTPγS. Active GTP-bound Ras was pulled down by GST-fusion Ras-binding domain of Raf1 (GST-Raf1-RBD) and detected by immunoblotting with Ras antibody. Lower panels: Total Ras is shown as the input control. C. Inhibition of <t>Rac1</t> GTPase by CRKL siRNA knockdown. Upper left panel: H3122 cell lysates were incubated with buffer (Ctrl), or as positive and negative controls, with GTPγS and GDP, respectively. Upper right panel: H3122 cell lysates with or without CRKL siRNA knockdown were incubated with GTPγS. Active GTP-bound Rac1 was pulled down by GST-fusion p21 binding domain of p21-activated kinase 1 (Pak1) (GST-Pak1-PBD) and detected by immunoblotting with Rac1 antibody. Lower panels: Total Rac1 serves as the input control. D. Cell viability assessed 72h after treatment with or without CRKL siRNAs by MTS assay, E. Cell migration, assessed using quantitative Boyden Chamber technique (16h after plating), and F. Colony formation assays (10 days after plating) were also performed in H3122 and H2228 cells with/without CRKL siRNA knockdown. Data represent the means of at least three independent experiments and are presented as percentage of untreated cells (Student's t -test: p -value
    Gtp Rac1 Pull Down Assay, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gtp rac1 pull down assay/product/Millipore
    Average 90 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    gtp rac1 pull down assay - by Bioz Stars, 2020-09
    90/100 stars
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    Effect of AA and SDS on the interaction of the p67 phox ·Rac-GTP complex with Nox2. A , SDS-PAGE analysis of purified proteins used in this study. GST alone, GST-Nox2-C, the wild-type p67 phox (1–212)-Rac1 (Q61L) chimera, and a mutant p67

    Journal: The Journal of Biological Chemistry

    Article Title: Arachidonic Acid Induces Direct Interaction of the p67phox-Rac Complex with the Phagocyte Oxidase Nox2, Leading to Superoxide Production *

    doi: 10.1074/jbc.M114.581785

    Figure Lengend Snippet: Effect of AA and SDS on the interaction of the p67 phox ·Rac-GTP complex with Nox2. A , SDS-PAGE analysis of purified proteins used in this study. GST alone, GST-Nox2-C, the wild-type p67 phox (1–212)-Rac1 (Q61L) chimera, and a mutant p67

    Article Snippet: For estimation of protein levels of FLAG-p47phox , Myc-p67phox , Myc-Rac1, and p22phox , proteins in cell lysates were subjected to SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore), and probed with an anti-FLAG monoclonal antibody (Sigma-Aldrich), an anti-Myc monoclonal antibody (Roche Applied Science), and an anti-p22phox polyclonal antibody (Santa Cruz Biotechnology), respectively.

    Techniques: SDS Page, Purification, Mutagenesis

    Role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with Rac. A , SDS-PAGE analysis of purified GST fusion proteins that were used in a GST pull-down assay for interaction with Rac. GST alone and GST fusion protein of p67 phox -(1–301) (the wild-type ( wt ) or a mutant protein carrying the V204A, Y198A, L199A, Y198A/V204A, L199A/V204A, or R102E substitution) was subjected to SDS-PAGE, followed by staining with Coomassie Brilliant Blue. The positions for marker proteins are indicated in kilodaltons. B , role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with Rac. GST-p67 phox proteins (the wild-type and indicated mutant proteins) were incubated with Rac1 (Q61L) and pulled down with glutathione-Sepharose 4B beads. The precipitated proteins were analyzed by immunoblot with the anti-Rac antibody, as described under “Experimental Procedures.” The data are representative of results from four independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: A Conserved Region between the TPR and Activation Domains of p67phox Participates in Activation of the Phagocyte NADPH Oxidase *

    doi: 10.1074/jbc.M110.161166

    Figure Lengend Snippet: Role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with Rac. A , SDS-PAGE analysis of purified GST fusion proteins that were used in a GST pull-down assay for interaction with Rac. GST alone and GST fusion protein of p67 phox -(1–301) (the wild-type ( wt ) or a mutant protein carrying the V204A, Y198A, L199A, Y198A/V204A, L199A/V204A, or R102E substitution) was subjected to SDS-PAGE, followed by staining with Coomassie Brilliant Blue. The positions for marker proteins are indicated in kilodaltons. B , role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with Rac. GST-p67 phox proteins (the wild-type and indicated mutant proteins) were incubated with Rac1 (Q61L) and pulled down with glutathione-Sepharose 4B beads. The precipitated proteins were analyzed by immunoblot with the anti-Rac antibody, as described under “Experimental Procedures.” The data are representative of results from four independent experiments.

    Article Snippet: For estimation of protein levels of HA-p67phox , FLAG-p47phox , Myc-Rac1, and p22phox , proteins in cell lysates were subjected to SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore), and probed with an anti-HA monoclonal antibody (Roche Applied Science), an anti-FLAG monoclonal antibody (Sigma-Aldrich), an anti-Myc monoclonal antibody (Roche Applied Science), and anti-p22phox polyclonal antibody (Santa Cruz Biotechnology), respectively.

    Techniques: SDS Page, Purification, Pull Down Assay, Mutagenesis, Staining, Marker, Incubation

    Role for the Nox activation region of p67 phox (amino acids 190–210) in gp91 phox /Nox2 activation. A , role for a region C-terminal to the TPR domain of p67 phox in a whole cell activation system of the phagocyte NADPH oxidase gp91 phox /Nox2. HA-tagged p67 phox (the wild-type ( wt ) or indicated mutant protein), FLAG-p47 phox , gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. B , role for Tyr-198, Leu-199, and Val-204 in activation of gp91 phox /Nox2. HA-tagged p67 phox carrying substitution for Tyr-198, Leu-199, or Val-204 as well as FLAG-p47 phox , gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. C , role for Tyr-198, Leu-199, and Val-204 in activation of gp91 phox /Nox2 in the presence of Rac1 (Q61L). HA-tagged p67 phox carrying the substitution for Tyr-198, Leu-199, or Val-204 as well as FLAG-p47 phox , Myc-Rac1 (Q61L), gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. Note that although the cDNA for Rac1 (Q61L) was transfected in C , neither wild-type nor mutant Rac1 was ectopically expressed in A and B . The protein levels of HA-p67 phox (the wild-type or indicated mutant protein), FLAG-p47 phox , Myc-Rac1 (Q61L), and p22 phox were analyzed by immunoblot with the anti-HA, anti-FLAG, anti-Myc, and anti-p22 phox antibodies, respectively, as described under “Experimental Procedures.” The transfected cells were incubated for 5 min at 37 °C and then stimulated with phorbol 12-myristate 13-acetate (200 ng/ml). The chemiluminescence change by the superoxide produced was continuously monitored with DIOGENES, as described under “Experimental Procedures.” Each graph represents the means ± S.D. of the chemiluminescence values integrated for 10 min, which were obtained from three independent transfections.

    Journal: The Journal of Biological Chemistry

    Article Title: A Conserved Region between the TPR and Activation Domains of p67phox Participates in Activation of the Phagocyte NADPH Oxidase *

    doi: 10.1074/jbc.M110.161166

    Figure Lengend Snippet: Role for the Nox activation region of p67 phox (amino acids 190–210) in gp91 phox /Nox2 activation. A , role for a region C-terminal to the TPR domain of p67 phox in a whole cell activation system of the phagocyte NADPH oxidase gp91 phox /Nox2. HA-tagged p67 phox (the wild-type ( wt ) or indicated mutant protein), FLAG-p47 phox , gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. B , role for Tyr-198, Leu-199, and Val-204 in activation of gp91 phox /Nox2. HA-tagged p67 phox carrying substitution for Tyr-198, Leu-199, or Val-204 as well as FLAG-p47 phox , gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. C , role for Tyr-198, Leu-199, and Val-204 in activation of gp91 phox /Nox2 in the presence of Rac1 (Q61L). HA-tagged p67 phox carrying the substitution for Tyr-198, Leu-199, or Val-204 as well as FLAG-p47 phox , Myc-Rac1 (Q61L), gp91 phox /Nox2, and p22 phox were co-expressed in CHO cells. Note that although the cDNA for Rac1 (Q61L) was transfected in C , neither wild-type nor mutant Rac1 was ectopically expressed in A and B . The protein levels of HA-p67 phox (the wild-type or indicated mutant protein), FLAG-p47 phox , Myc-Rac1 (Q61L), and p22 phox were analyzed by immunoblot with the anti-HA, anti-FLAG, anti-Myc, and anti-p22 phox antibodies, respectively, as described under “Experimental Procedures.” The transfected cells were incubated for 5 min at 37 °C and then stimulated with phorbol 12-myristate 13-acetate (200 ng/ml). The chemiluminescence change by the superoxide produced was continuously monitored with DIOGENES, as described under “Experimental Procedures.” Each graph represents the means ± S.D. of the chemiluminescence values integrated for 10 min, which were obtained from three independent transfections.

    Article Snippet: For estimation of protein levels of HA-p67phox , FLAG-p47phox , Myc-Rac1, and p22phox , proteins in cell lysates were subjected to SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore), and probed with an anti-HA monoclonal antibody (Roche Applied Science), an anti-FLAG monoclonal antibody (Sigma-Aldrich), an anti-Myc monoclonal antibody (Roche Applied Science), and anti-p22phox polyclonal antibody (Santa Cruz Biotechnology), respectively.

    Techniques: Activation Assay, Mutagenesis, Transfection, Incubation, Produced

    Role for the Nox activation region of p67 phox and the corresponding region of Noxa1 in Nox1 activation. A , activation of Nox1 by wild-type ( wt ) or mutant Noxa1 and Noxo1. HA-Noxa1 (the wild-type or indicated mutant protein), FLAG-Noxo1, Nox1, and p22 phox were co-expressed in CHO cells. B , activation of Nox1 by wild-type or mutant p67 phox and Noxo1 in the presence of Rac1 (Q61L). HA-p67 phox (the wild-type or indicated mutant protein), FLAG-Noxo1, Myc-Rac1 (Q61L), Nox1, and p22 phox were co-expressed in CHO cells. Protein levels of HA-p67 phox or HA-Noxa1, FLAG-Noxo1, Myc-Rac1 (Q61L), and p22 phox were analyzed by immunoblot with the anti-HA, anti-FLAG, anti-Myc, and anti-p22 phox antibodies, respectively, as described under “Experimental Procedures.” After preincubation for 5 min, the transfected cells were incubated at 37 °C with or without phorbol 12-myristate 13-acetate ( PMA , 200 ng/ml). Chemiluminescence change by superoxide produced was continuously monitored with DIOGENES, as described under “Experimental Procedures.” Each graph represents the means ± S.D. of the chemiluminescence values integrated for 10 min, which were obtained from three independent transfections.

    Journal: The Journal of Biological Chemistry

    Article Title: A Conserved Region between the TPR and Activation Domains of p67phox Participates in Activation of the Phagocyte NADPH Oxidase *

    doi: 10.1074/jbc.M110.161166

    Figure Lengend Snippet: Role for the Nox activation region of p67 phox and the corresponding region of Noxa1 in Nox1 activation. A , activation of Nox1 by wild-type ( wt ) or mutant Noxa1 and Noxo1. HA-Noxa1 (the wild-type or indicated mutant protein), FLAG-Noxo1, Nox1, and p22 phox were co-expressed in CHO cells. B , activation of Nox1 by wild-type or mutant p67 phox and Noxo1 in the presence of Rac1 (Q61L). HA-p67 phox (the wild-type or indicated mutant protein), FLAG-Noxo1, Myc-Rac1 (Q61L), Nox1, and p22 phox were co-expressed in CHO cells. Protein levels of HA-p67 phox or HA-Noxa1, FLAG-Noxo1, Myc-Rac1 (Q61L), and p22 phox were analyzed by immunoblot with the anti-HA, anti-FLAG, anti-Myc, and anti-p22 phox antibodies, respectively, as described under “Experimental Procedures.” After preincubation for 5 min, the transfected cells were incubated at 37 °C with or without phorbol 12-myristate 13-acetate ( PMA , 200 ng/ml). Chemiluminescence change by superoxide produced was continuously monitored with DIOGENES, as described under “Experimental Procedures.” Each graph represents the means ± S.D. of the chemiluminescence values integrated for 10 min, which were obtained from three independent transfections.

    Article Snippet: For estimation of protein levels of HA-p67phox , FLAG-p47phox , Myc-Rac1, and p22phox , proteins in cell lysates were subjected to SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore), and probed with an anti-HA monoclonal antibody (Roche Applied Science), an anti-FLAG monoclonal antibody (Sigma-Aldrich), an anti-Myc monoclonal antibody (Roche Applied Science), and anti-p22phox polyclonal antibody (Santa Cruz Biotechnology), respectively.

    Techniques: Activation Assay, Mutagenesis, Transfection, Incubation, Produced

    Role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with gp91 phox . A , SDS-PAGE analysis of purified, GST-tagged, or FLAG-tagged proteins that were used for interaction of p67 phox with gp91 phox . GST alone, GST-fused protein of gp91 phox -C, FLAG-tagged p67 phox (the wild-type ( wt ) or a mutant protein carrying the V204A, Y198A, L199A, Y198A/V204A, or L199A/V204A substitution), and Rac1 (Q61L) without a tag were subjected to SDS-PAGE, followed by staining with Coomassie Brilliant Blue. The positions for marker proteins are indicated in kilodaltons. B , role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with gp91 phox . GST-gp91 phox -C was incubated with FLAG-p67 phox proteins (the wild-type and indicated mutant proteins) in the presence of Rac1 (Q61L) and pulled down with glutathione-Sepharose 4B beads. The precipitated proteins were analyzed by immunoblot with the anti-FLAG antibody, as described under “Experimental Procedures.” The data are representative of results from four independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: A Conserved Region between the TPR and Activation Domains of p67phox Participates in Activation of the Phagocyte NADPH Oxidase *

    doi: 10.1074/jbc.M110.161166

    Figure Lengend Snippet: Role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with gp91 phox . A , SDS-PAGE analysis of purified, GST-tagged, or FLAG-tagged proteins that were used for interaction of p67 phox with gp91 phox . GST alone, GST-fused protein of gp91 phox -C, FLAG-tagged p67 phox (the wild-type ( wt ) or a mutant protein carrying the V204A, Y198A, L199A, Y198A/V204A, or L199A/V204A substitution), and Rac1 (Q61L) without a tag were subjected to SDS-PAGE, followed by staining with Coomassie Brilliant Blue. The positions for marker proteins are indicated in kilodaltons. B , role for Tyr-198, Leu-199, and Val-204 of p67 phox in interaction with gp91 phox . GST-gp91 phox -C was incubated with FLAG-p67 phox proteins (the wild-type and indicated mutant proteins) in the presence of Rac1 (Q61L) and pulled down with glutathione-Sepharose 4B beads. The precipitated proteins were analyzed by immunoblot with the anti-FLAG antibody, as described under “Experimental Procedures.” The data are representative of results from four independent experiments.

    Article Snippet: For estimation of protein levels of HA-p67phox , FLAG-p47phox , Myc-Rac1, and p22phox , proteins in cell lysates were subjected to SDS-PAGE, transferred to a polyvinylidene difluoride membrane (Millipore), and probed with an anti-HA monoclonal antibody (Roche Applied Science), an anti-FLAG monoclonal antibody (Sigma-Aldrich), an anti-Myc monoclonal antibody (Roche Applied Science), and anti-p22phox polyclonal antibody (Santa Cruz Biotechnology), respectively.

    Techniques: SDS Page, Purification, Mutagenesis, Staining, Marker, Incubation

    CRKL mediates ALK signaling and regulates cytoskeleton, cell migration and survival A. CRKL siRNA knockdown in H2228 and H3122. Protein lysates were extracted from cells treated with four individual CRLK siRNAs (20 nM) or their smartpool for 72 h, and subjected to Western blot analyses. The graph shows the quantification of CRKL proteins. B. Inhibition of Ras GTPase activity by CRKL siRNA knockdown. Upper left panel: Lysates of untreated H3122 cells were incubated with buffer (Ctrl), non-hydrolyzable analog of GTP (GTPγS as a positive control) or GDP (as a negative control). Upper right panel: Lysates of H3122 cells treated with or without CRKL siRNA were incubated with GTPγS. Active GTP-bound Ras was pulled down by GST-fusion Ras-binding domain of Raf1 (GST-Raf1-RBD) and detected by immunoblotting with Ras antibody. Lower panels: Total Ras is shown as the input control. C. Inhibition of Rac1 GTPase by CRKL siRNA knockdown. Upper left panel: H3122 cell lysates were incubated with buffer (Ctrl), or as positive and negative controls, with GTPγS and GDP, respectively. Upper right panel: H3122 cell lysates with or without CRKL siRNA knockdown were incubated with GTPγS. Active GTP-bound Rac1 was pulled down by GST-fusion p21 binding domain of p21-activated kinase 1 (Pak1) (GST-Pak1-PBD) and detected by immunoblotting with Rac1 antibody. Lower panels: Total Rac1 serves as the input control. D. Cell viability assessed 72h after treatment with or without CRKL siRNAs by MTS assay, E. Cell migration, assessed using quantitative Boyden Chamber technique (16h after plating), and F. Colony formation assays (10 days after plating) were also performed in H3122 and H2228 cells with/without CRKL siRNA knockdown. Data represent the means of at least three independent experiments and are presented as percentage of untreated cells (Student's t -test: p -value

    Journal: Oncotarget

    Article Title: CRKL mediates EML4-ALK signaling and is a potential therapeutic target for ALK-rearranged lung adenocarcinoma

    doi: 10.18632/oncotarget.8638

    Figure Lengend Snippet: CRKL mediates ALK signaling and regulates cytoskeleton, cell migration and survival A. CRKL siRNA knockdown in H2228 and H3122. Protein lysates were extracted from cells treated with four individual CRLK siRNAs (20 nM) or their smartpool for 72 h, and subjected to Western blot analyses. The graph shows the quantification of CRKL proteins. B. Inhibition of Ras GTPase activity by CRKL siRNA knockdown. Upper left panel: Lysates of untreated H3122 cells were incubated with buffer (Ctrl), non-hydrolyzable analog of GTP (GTPγS as a positive control) or GDP (as a negative control). Upper right panel: Lysates of H3122 cells treated with or without CRKL siRNA were incubated with GTPγS. Active GTP-bound Ras was pulled down by GST-fusion Ras-binding domain of Raf1 (GST-Raf1-RBD) and detected by immunoblotting with Ras antibody. Lower panels: Total Ras is shown as the input control. C. Inhibition of Rac1 GTPase by CRKL siRNA knockdown. Upper left panel: H3122 cell lysates were incubated with buffer (Ctrl), or as positive and negative controls, with GTPγS and GDP, respectively. Upper right panel: H3122 cell lysates with or without CRKL siRNA knockdown were incubated with GTPγS. Active GTP-bound Rac1 was pulled down by GST-fusion p21 binding domain of p21-activated kinase 1 (Pak1) (GST-Pak1-PBD) and detected by immunoblotting with Rac1 antibody. Lower panels: Total Rac1 serves as the input control. D. Cell viability assessed 72h after treatment with or without CRKL siRNAs by MTS assay, E. Cell migration, assessed using quantitative Boyden Chamber technique (16h after plating), and F. Colony formation assays (10 days after plating) were also performed in H3122 and H2228 cells with/without CRKL siRNA knockdown. Data represent the means of at least three independent experiments and are presented as percentage of untreated cells (Student's t -test: p -value

    Article Snippet: GTP-RAS and GTP-RAC1 pull-down assay The pull-down of GTP-bound RAS and RAC1 was performed by the use of active RAS and RAC1 pull-down and detection kits (Millipore) respectively according to the manufacturer's instructions.

    Techniques: Migration, Western Blot, Inhibition, Activity Assay, Incubation, Positive Control, Negative Control, Binding Assay, MTS Assay

    ARV-activated Rac1 through activation of p38 MAPK and Src. DF-1 cells were grown in 6-cm 2 cell culture dishes to 75% confluence, and then the cells were cultured in serum-free media overnight. Pulldown assay for GTP-Rac1 was performed. A , DF-1 cells were

    Journal: The Journal of Biological Chemistry

    Article Title: Cell Entry of Avian Reovirus Follows a Caveolin-1-mediated and Dynamin-2-dependent Endocytic Pathway That Requires Activation of p38 Mitogen-activated Protein Kinase (MAPK) and Src Signaling Pathways as Well as Microtubules and Small GTPase Rab5 Protein *

    doi: 10.1074/jbc.M111.257154

    Figure Lengend Snippet: ARV-activated Rac1 through activation of p38 MAPK and Src. DF-1 cells were grown in 6-cm 2 cell culture dishes to 75% confluence, and then the cells were cultured in serum-free media overnight. Pulldown assay for GTP-Rac1 was performed. A , DF-1 cells were

    Article Snippet: GTP-bound Rac1 was isolated from DF-1 cell lysates using a Rac1 activation assay kit (Millipore; San Diego) according to the manufacturer's protocol.

    Techniques: Activation Assay, Cell Culture