rnase ribonuclease inhibitor  (Millipore)


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    Name:
    Ribonuclease Inhibitor
    Description:
    Ribonuclease inhibitor works to inhibit RNase activity by forming a tight non covalent 1 1 complex It is derived from E coli that express portions of the human placental ribonuclease inhibitor It inhibits RNases A B and C It will not inhibit RNases H 1 T1 S1 Nuclease SP6 T7 RNA Polymerase T3 RNA Polymerase AMV Reverse Transcriptase M MLV Reverse Transcriptase or Taq Polymerase The inhibitor can be removed by phenol extraction or by heating to 65C for 10 minutes
    Catalog Number:
    r1158
    Price:
    None
    Applications:
    Useful for in vitro inhibition of ribonucleases, including procedures like cDNA synthesis, RT-PCR, and in vitro transcription and translation.
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    Structured Review

    Millipore rnase ribonuclease inhibitor
    Ribonuclease inhibitor works to inhibit RNase activity by forming a tight non covalent 1 1 complex It is derived from E coli that express portions of the human placental ribonuclease inhibitor It inhibits RNases A B and C It will not inhibit RNases H 1 T1 S1 Nuclease SP6 T7 RNA Polymerase T3 RNA Polymerase AMV Reverse Transcriptase M MLV Reverse Transcriptase or Taq Polymerase The inhibitor can be removed by phenol extraction or by heating to 65C for 10 minutes
    https://www.bioz.com/result/rnase ribonuclease inhibitor/product/Millipore
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rnase ribonuclease inhibitor - by Bioz Stars, 2020-11
    99/100 stars

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    Related Articles

    Transfection:

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

    Protease Inhibitor:

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

    RNA Extraction:

    Article Title: Esf2p, a U3-Associated Factor Required for Small-Subunit Processome Assembly and Compaction
    Article Snippet: .. For the depletion of Esf2p protein, the tet :: esf2 strain (and the isogenic wild-type control strain R1158) was exposed to 10 μg/ml doxycycline (DOX) (Sigma) for a total of up to 24 h before harvesting for RNA extraction. ..

    Purification:

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

    Immunoprecipitation:

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

    Incubation:

    Article Title: N6-methyladenosine alters RNA structure to regulate binding of a low-complexity protein
    Article Snippet: .. The HNRNPG PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1× IP buffer (50 mM Tris-Cl (pH 7.4), 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4°C for 2 h under gentle shaking conditions. .. For each PAR-CLIP–MeRIP experiment, 20 μl Protein A beads (10002D, Thermo Scientific) were washed twice with 1 ml 1× IP buffer, blocked by a 2-h incubation with 100 μl 1× IP buffer supplemented with BSA (0.5 mg/ml), RNasin and human placental RNase inhibitor, and then washed twice with 100 μl 1× IP buffer.

    Article Title: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions
    Article Snippet: .. The hnRNP C PAR-CLIP RNA sample was incubated with m6 A-specific antibody (202003, SYSY), RNase inhibitor (80 units, Sigma-Aldrich), human placental RNase inhibitor (NEB) in 200 μl 1x IP buffer (50 mM Tris-HCl pH 7.4, 750 mM NaCl and 0.5% (vol/vol) Igepal CA-630) at 4 °C for 2 hours under gentle shaking conditions. .. For each PARCLIP-MeRIP experiment, 20 μl protein-A beads (Invitrogen) were washed twice with 1 ml 1x IP buffer, blocked with 2 hours incubation with 100 μl 1× IP buffer supplemented with BSA (0.5 mg/ml), RNasin and Human placental RNase inhibitor, and then washed twice with 100 μl 1x IP buffer.

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

    Fractionation:

    Article Title: Integrative genomics positions MKRN1 as a novel ribonucleoprotein within the embryonic stem cell gene regulatory network
    Article Snippet: .. Fractionation of cellular compartments A total of 108 MKRN1 knockdown (shMKRN1) or shGFP control ESCs were resuspended in PBS, washed (500 g , 5 min) with 5 ml 0.25 M sucrose solution pH 7.2, and resuspended in 1 ml 0.25 M sucrose containing ribonuclease inhibitor (1 mM ribonucleoside vanadyl, Sigma‐Aldrich). ..

    Negative Control:

    Article Title: Inhibition of the MID1 protein complex: a novel approach targeting APP protein synthesis
    Article Snippet: .. 48 h after transfection, cells were treated with or without 2.5 mM metformin and incubated another 24 h. After UV-crosslinking (200 mJ/cm2 ) cells were lysed in TKM buffer (20 mM Tris pH 7.4, 100 mM KCl, 5 mM MgCl2 , Complete protease inhibitor cocktail (Roche), RNAse inhibitor, 0.2% NP40) and MID1 protein complexes were purified by immunoprecipitation using anti-FLAG M1 Agarose Affinity Gel (Sigma-Aldrich) or IgG-agarose (Sigma-Aldrich) as a negative control. .. Protein-bound mRNA was isolated after DNAse and proteinase K digestion by phenol-chloroform purification and analyzed by RT-PCR.

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    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 1 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 1 article reviews
    Price from $9.99 to $1999.99
    gtp rac1 pull down assay - by Bioz Stars, 2020-11
    90/100 stars
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    96
    Millipore rac1
    A : Bar graph shows mean ± SEM of <t>Rac1</t> knockout efficiency in soleus and EDL muscle of muscle-specific inducible Rac1 KO mice and WT controls ( n = 11–13). B : Representative Western blots showing the effect of doxycycline (1 g/L) treatment on Rac1 protein in WT vs. Floxed Rac1 KO mice (Rac1 fl/fl*Cre) (gastrocnemius muscle). C : Contraction-stimulated 2DG uptake in incubated isolated soleus and EDL muscles from WT and Rac1 KO mice ( n = 11–13). D : Contraction-stimulated fold changes from basal in 2DG uptake in soleus and EDL from WT and Rac1 KO mice. E : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 , p-TBC1D4 Thr642 (Soleus), p-TBC1D1 Ser237 (EDL), and actin, GLUT4, and Rac1 total proteins in response to contraction. F : Bar graph shows mean ± SEM of contraction-induced p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 /PAK1, p-TBC1D4 Thr542 , and p-TBC1D1 Ser237 in soleus and EDL from Rac1 KO mice and WT controls. G : Bar graph shows mean ± SEM of total PAK1 and GLUT4 protein in soleus and EDL of WT and Rac1 KO mice. Statistical significances between basal and contraction are indicated by * P
    Rac1, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rac1/product/Millipore
    Average 96 stars, based on 30 article reviews
    Price from $9.99 to $1999.99
    rac1 - by Bioz Stars, 2020-11
    96/100 stars
      Buy from Supplier

    Image Search Results


    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

    A : Bar graph shows mean ± SEM of Rac1 knockout efficiency in soleus and EDL muscle of muscle-specific inducible Rac1 KO mice and WT controls ( n = 11–13). B : Representative Western blots showing the effect of doxycycline (1 g/L) treatment on Rac1 protein in WT vs. Floxed Rac1 KO mice (Rac1 fl/fl*Cre) (gastrocnemius muscle). C : Contraction-stimulated 2DG uptake in incubated isolated soleus and EDL muscles from WT and Rac1 KO mice ( n = 11–13). D : Contraction-stimulated fold changes from basal in 2DG uptake in soleus and EDL from WT and Rac1 KO mice. E : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 , p-TBC1D4 Thr642 (Soleus), p-TBC1D1 Ser237 (EDL), and actin, GLUT4, and Rac1 total proteins in response to contraction. F : Bar graph shows mean ± SEM of contraction-induced p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 /PAK1, p-TBC1D4 Thr542 , and p-TBC1D1 Ser237 in soleus and EDL from Rac1 KO mice and WT controls. G : Bar graph shows mean ± SEM of total PAK1 and GLUT4 protein in soleus and EDL of WT and Rac1 KO mice. Statistical significances between basal and contraction are indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Bar graph shows mean ± SEM of Rac1 knockout efficiency in soleus and EDL muscle of muscle-specific inducible Rac1 KO mice and WT controls ( n = 11–13). B : Representative Western blots showing the effect of doxycycline (1 g/L) treatment on Rac1 protein in WT vs. Floxed Rac1 KO mice (Rac1 fl/fl*Cre) (gastrocnemius muscle). C : Contraction-stimulated 2DG uptake in incubated isolated soleus and EDL muscles from WT and Rac1 KO mice ( n = 11–13). D : Contraction-stimulated fold changes from basal in 2DG uptake in soleus and EDL from WT and Rac1 KO mice. E : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 , p-TBC1D4 Thr642 (Soleus), p-TBC1D1 Ser237 (EDL), and actin, GLUT4, and Rac1 total proteins in response to contraction. F : Bar graph shows mean ± SEM of contraction-induced p-AMPK Thr172 , p-ACC Ser212 , PAS, p-PAK Thr423 /PAK1, p-TBC1D4 Thr542 , and p-TBC1D1 Ser237 in soleus and EDL from Rac1 KO mice and WT controls. G : Bar graph shows mean ± SEM of total PAK1 and GLUT4 protein in soleus and EDL of WT and Rac1 KO mice. Statistical significances between basal and contraction are indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Knock-Out, Mouse Assay, Western Blot, Incubation, Isolation

    A : Contraction-stimulated 2DG uptake in soleus and EDL without or with 200 μmol/L NSC23766, 1-h preincubation ( n = 12). B : Contraction-stimulated 2DG uptake in soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 10). C : Representative blots and bar graphs show contraction-stimulated p-AMPK Thr172 and p-ACC Ser212 in soleus and EDL muscle without or with 200 μmol/L NSC23766. D : Representative blots and bar graphs show contraction-stimulated p-AMPK Thr172 , p-ACC Ser212 , and p-PAK Thr423 signaling and Rac1 and GLUT4 total protein in soleus and EDL muscle without or with 10 μmol/L Rac1 Inhibitor II (Inhib II). E : Insulin-stimulated (60 nmol/L) 2DG uptake in mouse white adipose (gonadal) tissue without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 8). Statistical significance between basal and contraction is indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Contraction-stimulated 2DG uptake in soleus and EDL without or with 200 μmol/L NSC23766, 1-h preincubation ( n = 12). B : Contraction-stimulated 2DG uptake in soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 10). C : Representative blots and bar graphs show contraction-stimulated p-AMPK Thr172 and p-ACC Ser212 in soleus and EDL muscle without or with 200 μmol/L NSC23766. D : Representative blots and bar graphs show contraction-stimulated p-AMPK Thr172 , p-ACC Ser212 , and p-PAK Thr423 signaling and Rac1 and GLUT4 total protein in soleus and EDL muscle without or with 10 μmol/L Rac1 Inhibitor II (Inhib II). E : Insulin-stimulated (60 nmol/L) 2DG uptake in mouse white adipose (gonadal) tissue without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 8). Statistical significance between basal and contraction is indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Inhibition

    A : Rac1-GTP binding in incubated mouse soleus muscle stimulated with (black bar) or without (white bar) electrically induced contraction (100 Hz 15-s intervals, 2-s train, 0.2-ms impulses) ( n = 7). B : Rac1-GTP binding in incubated mouse soleus muscle kept at resting tension (2–5 mN; basal) or stretched (150 mN, 15 min) ( n = 7). C : Mouse quadriceps muscle, basal and after 30 min running at speed of 10 m/min (exercise 10 m/min = 40% of max running speed) ( n = 7). D : Rac1-GTP binding ( top ) and p-PAK Thr423 ( middle ), and representative blots ( bottom ) in mouse quadriceps muscle basal or in response to 30-min treadmill running at 50 or 70% (Ex 50/70%) of maximum running speed ( n = 8). E : Representative images and bar graph (FWHM) showing Rac1 staining of single fibers isolated from mouse EDL muscle in the basal state or after 30 min of treadmill running at 70% of maximum running speed ( n = 5). Statistical significance compared with basal is indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Rac1-GTP binding in incubated mouse soleus muscle stimulated with (black bar) or without (white bar) electrically induced contraction (100 Hz 15-s intervals, 2-s train, 0.2-ms impulses) ( n = 7). B : Rac1-GTP binding in incubated mouse soleus muscle kept at resting tension (2–5 mN; basal) or stretched (150 mN, 15 min) ( n = 7). C : Mouse quadriceps muscle, basal and after 30 min running at speed of 10 m/min (exercise 10 m/min = 40% of max running speed) ( n = 7). D : Rac1-GTP binding ( top ) and p-PAK Thr423 ( middle ), and representative blots ( bottom ) in mouse quadriceps muscle basal or in response to 30-min treadmill running at 50 or 70% (Ex 50/70%) of maximum running speed ( n = 8). E : Representative images and bar graph (FWHM) showing Rac1 staining of single fibers isolated from mouse EDL muscle in the basal state or after 30 min of treadmill running at 70% of maximum running speed ( n = 5). Statistical significance compared with basal is indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Binding Assay, Incubation, Mass Spectrometry, Staining, Isolation

    A : Rac1 protein expression in mouse soleus (SOL), extensor digitorum longus (EDL), and gastrocnemius (Gast) ( n = 9). B : Rac1 expression in human soleus and gastrocnemius ( n = 8). C : Intracellular localization of Rac1 and VAMP3 in mouse hind limb (gastrocnemius and quadriceps) muscle. L, lysate; P1 and 2, plasma membrane fractions; Cyt, cytosol; F1-8 intracellular fractions ( n = 7). Statistical significance is indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Rac1 protein expression in mouse soleus (SOL), extensor digitorum longus (EDL), and gastrocnemius (Gast) ( n = 9). B : Rac1 expression in human soleus and gastrocnemius ( n = 8). C : Intracellular localization of Rac1 and VAMP3 in mouse hind limb (gastrocnemius and quadriceps) muscle. L, lysate; P1 and 2, plasma membrane fractions; Cyt, cytosol; F1-8 intracellular fractions ( n = 7). Statistical significance is indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Expressing

    A : Rac1-GTP binding in human soleus (SOL) and gastrocnemius (Gast) in the basal state and after 45 min of inclined walking at ∼69% V o 2 peak ( n = 5–9). B : Exercise-stimulated fold changes from basal in soleus (SOL) and gastrocnemius (Gast). C : Bar graph and representative Western blot analysis of p-Rac1 Ser71 in response to exercise in human soleus and gastrocnemius muscle ( n = 5–9). D : Bar graph and Western blot analysis of p-PAK Thr423 in response to exercise in human soleus and gastrocnemius muscle ( n = 5–9). Statistical significance compared with basal is indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Rac1-GTP binding in human soleus (SOL) and gastrocnemius (Gast) in the basal state and after 45 min of inclined walking at ∼69% V o 2 peak ( n = 5–9). B : Exercise-stimulated fold changes from basal in soleus (SOL) and gastrocnemius (Gast). C : Bar graph and representative Western blot analysis of p-Rac1 Ser71 in response to exercise in human soleus and gastrocnemius muscle ( n = 5–9). D : Bar graph and Western blot analysis of p-PAK Thr423 in response to exercise in human soleus and gastrocnemius muscle ( n = 5–9). Statistical significance compared with basal is indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Binding Assay, Western Blot

    A : Rac1-GTP binding after ex vivo AICAR (2 mmol/L) stimulation of mouse soleus and EDL muscles ( n = 5–6). B : Bar graphs and representative blots showing p-AMPK Thr172 and p-ACC Ser212 in response to AICAR ( n = 5–6). C : Representative blots and bar graphs of the effect of insulin (100 nmol/L; 10 min), DNP (0.5 mmol/L; 20 min), and AICAR (2 mmol/L; 30 min) on Rac1-GTP binding in C2C12 myotubes ( n = 5). D : Rac1-GTP binding in WT and AMPK KD mouse quadriceps muscle basal or in response to 30 min of treadmill running at 70% (Exe 70%) of the maximum running speed ( n = 6–9). Statistical significance compared with basal is indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : Rac1-GTP binding after ex vivo AICAR (2 mmol/L) stimulation of mouse soleus and EDL muscles ( n = 5–6). B : Bar graphs and representative blots showing p-AMPK Thr172 and p-ACC Ser212 in response to AICAR ( n = 5–6). C : Representative blots and bar graphs of the effect of insulin (100 nmol/L; 10 min), DNP (0.5 mmol/L; 20 min), and AICAR (2 mmol/L; 30 min) on Rac1-GTP binding in C2C12 myotubes ( n = 5). D : Rac1-GTP binding in WT and AMPK KD mouse quadriceps muscle basal or in response to 30 min of treadmill running at 70% (Exe 70%) of the maximum running speed ( n = 6–9). Statistical significance compared with basal is indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Binding Assay, Ex Vivo

    A : AICAR-stimulated (2 mmol/L) 2DG uptake in incubated isolated soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 8). B : Bar graphs show mean ± SEM of AICAR-induced p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , in soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II) ( n = 8). C : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , and GLUT4 and Rac1 total proteins in response to AICAR without or with 10 μmol/L Rac1 Inhibitor II (Inhib II). D : AICAR-stimulated 2DG uptake in mouse soleus and EDL muscles from WT and muscle-specific inducible Rac1 KO ( n = 8) mice. E : Bar graphs show mean ± SEM of AICAR-induced p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , and total Rac1 and PAK1 protein in soleus and EDL muscles from WT or Rac1 KO mice ( n = 8). F : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , and p-PAK Thr423 /PAK1, and GLUT4, Rac1, and PAK1 total proteins in response to AICAR in WT and Rac1 KO soleus and EDL muscles. G : AICAR-stimulated GLUT4 translocation in C2C12-GLUT4myc myotubes in response to DNP (0.5 mmol/L) or AICAR (2 mmol/L) without or with 200 μmol/L NSC23766, 1-h preincubation ( n = 6). Statistical significances between basal and AICAR are indicated by * P

    Journal: Diabetes

    Article Title: Rac1 Is a Novel Regulator of Contraction-Stimulated Glucose Uptake in Skeletal Muscle

    doi: 10.2337/db12-0491

    Figure Lengend Snippet: A : AICAR-stimulated (2 mmol/L) 2DG uptake in incubated isolated soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II), 1-h preincubation ( n = 8). B : Bar graphs show mean ± SEM of AICAR-induced p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , in soleus and EDL without or with 10 μmol/L Rac1 Inhibitor II (Inhib II) ( n = 8). C : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , and GLUT4 and Rac1 total proteins in response to AICAR without or with 10 μmol/L Rac1 Inhibitor II (Inhib II). D : AICAR-stimulated 2DG uptake in mouse soleus and EDL muscles from WT and muscle-specific inducible Rac1 KO ( n = 8) mice. E : Bar graphs show mean ± SEM of AICAR-induced p-AMPK Thr172 , p-ACC Ser212 , p-PAK Thr423 , and total Rac1 and PAK1 protein in soleus and EDL muscles from WT or Rac1 KO mice ( n = 8). F : Representative Western blots of p-AMPK Thr172 , p-ACC Ser212 , and p-PAK Thr423 /PAK1, and GLUT4, Rac1, and PAK1 total proteins in response to AICAR in WT and Rac1 KO soleus and EDL muscles. G : AICAR-stimulated GLUT4 translocation in C2C12-GLUT4myc myotubes in response to DNP (0.5 mmol/L) or AICAR (2 mmol/L) without or with 200 μmol/L NSC23766, 1-h preincubation ( n = 6). Statistical significances between basal and AICAR are indicated by * P

    Article Snippet: When interpreted together with the results using Rac1 inhibitors, these data strongly implicate Rac1 in the regulation of contraction-stimulated glucose uptake in skeletal muscle.

    Techniques: Incubation, Isolation, Inhibition, Western Blot, Mouse Assay, Translocation Assay

    Nuclear expression of Rac1 in squamous intraepithelial lesions and cervical cancer cell lines . A . Representative images showing nuclear Rac1 expression in low-grade squamous intraepithelial lesions (L-SIL) and high-grade squamous intraepithelial lesions (H-SIL) but not in epithelium without squamous SIL. B. Representative images of immunocytochemical analysis showing nuclear Rac1 expression in cervical cancer cells C33A and SiHa, but not in non-tumorigenic Hacat cells. Magnification: 40×. C. Western blot analysis of Rac1 protein levels in cytoplasmic (C) and nuclear (N) protein extracts from HaCat, C33A and SiHa cells. D. Western blot analysis of Rac1 and Tiam1 protein levels in whole-cell extracts from HaCat, C33A and SiHa cells.

    Journal: BMC Cancer

    Article Title: Nuclear expression of Rac1 in cervical premalignant lesions and cervical cancer cells

    doi: 10.1186/1471-2407-12-116

    Figure Lengend Snippet: Nuclear expression of Rac1 in squamous intraepithelial lesions and cervical cancer cell lines . A . Representative images showing nuclear Rac1 expression in low-grade squamous intraepithelial lesions (L-SIL) and high-grade squamous intraepithelial lesions (H-SIL) but not in epithelium without squamous SIL. B. Representative images of immunocytochemical analysis showing nuclear Rac1 expression in cervical cancer cells C33A and SiHa, but not in non-tumorigenic Hacat cells. Magnification: 40×. C. Western blot analysis of Rac1 protein levels in cytoplasmic (C) and nuclear (N) protein extracts from HaCat, C33A and SiHa cells. D. Western blot analysis of Rac1 and Tiam1 protein levels in whole-cell extracts from HaCat, C33A and SiHa cells.

    Article Snippet: Therefore, it is possible that inhibition of the cytoplasmic pool of Rac1 in both cervical cancer-derived and non-tumorigenic cells may result in a reduction of cell proliferation, independently of Rac1 nuclear functions.

    Techniques: Expressing, Western Blot

    Expression of Rho-GTPases and Rho-GEFs in cervical epithelium without squamous intraepithelial lesions, low-grade squamous intraepithelial lesions (L-SIL), and high-grade squamous intraepithelial lesions (H-SIL) . Representative images of immunohistochemical analysis of Rac1, RhoA, Cdc42, Tiam, and beta-Pix expression. Magnification: 40 ×.

    Journal: BMC Cancer

    Article Title: Nuclear expression of Rac1 in cervical premalignant lesions and cervical cancer cells

    doi: 10.1186/1471-2407-12-116

    Figure Lengend Snippet: Expression of Rho-GTPases and Rho-GEFs in cervical epithelium without squamous intraepithelial lesions, low-grade squamous intraepithelial lesions (L-SIL), and high-grade squamous intraepithelial lesions (H-SIL) . Representative images of immunohistochemical analysis of Rac1, RhoA, Cdc42, Tiam, and beta-Pix expression. Magnification: 40 ×.

    Article Snippet: Therefore, it is possible that inhibition of the cytoplasmic pool of Rac1 in both cervical cancer-derived and non-tumorigenic cells may result in a reduction of cell proliferation, independently of Rac1 nuclear functions.

    Techniques: Expressing, Immunohistochemistry

    Effect of the Rac1 chemical inhibitor NSC23677 on the subcellular localization of Rac1, and on cell proliferation . A . Representative images of immunocytochemical analysis for Rac1 in C33A and SiHa cells, treated or not with the Rac1 inhibitor NSC23677. B . Detection of Rac1 in cytoplasmic ( C ) and nuclear (N) proteins from HaCat, C33A and SiHa cells treated or not with the Rac1 inhibitor NSC23677. Lamin B was used as a nuclear marker and α-tubilin is a cytoplasmic marker. C . Results from crystal violet cell proliferation assays on HaCat, C33A and SiHa cells treated or not with the Rac1 inhibitor NSC23766 for 48 h. Data shown are average optical density values plus standard deviation of experiments performed in triplicate.

    Journal: BMC Cancer

    Article Title: Nuclear expression of Rac1 in cervical premalignant lesions and cervical cancer cells

    doi: 10.1186/1471-2407-12-116

    Figure Lengend Snippet: Effect of the Rac1 chemical inhibitor NSC23677 on the subcellular localization of Rac1, and on cell proliferation . A . Representative images of immunocytochemical analysis for Rac1 in C33A and SiHa cells, treated or not with the Rac1 inhibitor NSC23677. B . Detection of Rac1 in cytoplasmic ( C ) and nuclear (N) proteins from HaCat, C33A and SiHa cells treated or not with the Rac1 inhibitor NSC23677. Lamin B was used as a nuclear marker and α-tubilin is a cytoplasmic marker. C . Results from crystal violet cell proliferation assays on HaCat, C33A and SiHa cells treated or not with the Rac1 inhibitor NSC23766 for 48 h. Data shown are average optical density values plus standard deviation of experiments performed in triplicate.

    Article Snippet: Therefore, it is possible that inhibition of the cytoplasmic pool of Rac1 in both cervical cancer-derived and non-tumorigenic cells may result in a reduction of cell proliferation, independently of Rac1 nuclear functions.

    Techniques: Marker, Standard Deviation

    mDia1 regulates RAGE-induced c-Src translocation to the membrane and consequent Rac1 activation, ROS generation and cellular signaling in SMCs

    Journal: Circulation Research

    Article Title: The Formin mDia1 Mediates Vascular Remodeling via Integration of Oxidative Signal Transduction Pathways

    doi: 10.1161/CIRCRESAHA.111.262519

    Figure Lengend Snippet: mDia1 regulates RAGE-induced c-Src translocation to the membrane and consequent Rac1 activation, ROS generation and cellular signaling in SMCs

    Article Snippet: In human umbilical vein endothelial cells, exposure to thrombin resulted in rapid activation of RhoA and inhibition of rac1; in contrast, stimulation of these cells with the bioactive phospholipid sphingosine-1-phospate (S1P) resulted in very modest and delayed activation of RhoA with strong activation of rac1.

    Techniques: Translocation Assay, Activation Assay

    mDia1, Rac1 and Nox1 are involved in RAGE induced generation of ROS and consequent cellular signaling in SMCs

    Journal: Circulation Research

    Article Title: The Formin mDia1 Mediates Vascular Remodeling via Integration of Oxidative Signal Transduction Pathways

    doi: 10.1161/CIRCRESAHA.111.262519

    Figure Lengend Snippet: mDia1, Rac1 and Nox1 are involved in RAGE induced generation of ROS and consequent cellular signaling in SMCs

    Article Snippet: In human umbilical vein endothelial cells, exposure to thrombin resulted in rapid activation of RhoA and inhibition of rac1; in contrast, stimulation of these cells with the bioactive phospholipid sphingosine-1-phospate (S1P) resulted in very modest and delayed activation of RhoA with strong activation of rac1.

    Techniques: