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  • 86
    Roche collagenase dispase
    PTPN13 or β-Catenin Silencing Increases Cell Adhesiveness and the Upregulation of Cell Membrane Adhesion Molecules (A) The proportion of Lin – (CD45 + Lin – ) and LSKs (CD45 + Lin – c-KIT + SCA1 + ) cells was analyzed in <t>collagenase-/dispase-digested</t> bones after BM had been flushed out (n = 4–5 different mice). The frequency of Lin – and LSKs strongly attached to the bone was higher in mice in which PTPN13 or β-catenin had been silenced. (B and C) PTPN13 or β-catenin was silenced in HEL cells. The levels of expression were analyzed by qRT-PCR (n = 5 or 8 independent experiments) (B) and immunoblotting (n = 3 independent experiments) (C). (D) Cell adhesiveness to collagen- or fibronectin-coated plates was analyzed in these cells (n = 5–7 independent experiments). (E) The expression of several genes coding for CAMs was analyzed by qRT-PCR (n = 4–9 independent experiments). (F) CDH1 promoter activity was analyzed by luciferase reporter assay in the presence or absence of a β-catenin stable mutant (β-catenin S33Y) (n = 4 independent experiments). Means ± SD are shown. See also Figure S4 .
    Collagenase Dispase, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore collagenase dispase
    PTPN13 or β-Catenin Silencing Increases Cell Adhesiveness and the Upregulation of Cell Membrane Adhesion Molecules (A) The proportion of Lin – (CD45 + Lin – ) and LSKs (CD45 + Lin – c-KIT + SCA1 + ) cells was analyzed in <t>collagenase-/dispase-digested</t> bones after BM had been flushed out (n = 4–5 different mice). The frequency of Lin – and LSKs strongly attached to the bone was higher in mice in which PTPN13 or β-catenin had been silenced. (B and C) PTPN13 or β-catenin was silenced in HEL cells. The levels of expression were analyzed by qRT-PCR (n = 5 or 8 independent experiments) (B) and immunoblotting (n = 3 independent experiments) (C). (D) Cell adhesiveness to collagen- or fibronectin-coated plates was analyzed in these cells (n = 5–7 independent experiments). (E) The expression of several genes coding for CAMs was analyzed by qRT-PCR (n = 4–9 independent experiments). (F) CDH1 promoter activity was analyzed by luciferase reporter assay in the presence or absence of a β-catenin stable mutant (β-catenin S33Y) (n = 4 independent experiments). Means ± SD are shown. See also Figure S4 .
    Collagenase Dispase, supplied by Millipore, 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/collagenase dispase/product/Millipore
    Average 99 stars, based on 1 article reviews
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    86
    Roche collagenase dispase solution
    Characterization of module thrombogenicity using whole-blood studies. ( a ) Clot formation times. The presence of HUVEC on the modules significantly increased the time to clot formation ( P = 1.4 × 10 −5 ) of slightly heparinized whole blood (0.75 units/ml) in a clotting test. In some cases, clot formation never actually occurred, and the test was terminated between 4,500 and 5,400 s; in these instances, the recorded time was the test termination time. Mean clot time is represented by the thick central line within the box. Open circles and stars represent outliers and extreme outliers, respectively. ( b ) Fresh whole blood (0.75 units/ml heparin) perfused through a HUVEC-covered modular construct (filled circles) maintains platelet levels no different from those measured in the absence of modules (open circles, flow circuit blank; includes polypropylene mesh required to keep modules in place). Blood perfusion through a control modular construct in which HUVEC have been removed by <t>dispase–collagenase</t> action (open squares), however, results in significant reductions in platelet number, indicating platelet activation and the thrombogenic response that occurs in the absence of HUVEC. Error bars indicate SEM ( n = 3, 4, and 7 for background, dispase-treated modular constructs, and HUVEC-covered modular constructs, respectively).
    Collagenase Dispase Solution, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/collagenase dispase solution/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase dispase solution - by Bioz Stars, 2021-03
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    86
    Roche dispase
    Overview of retinal endothelial cell isolation for RNA sequencing P5 retinas from wild‐type mice were digested with collagenase and <t>dispase.</t> CD31 and CD34 double‐positive endothelial cells were isolated by FACS, pooled into two samples, and analyzed by RNA‐Seq on an Illumina HiSeq 4000 sequencer generating 50 bp single‐end reads ( ca. 30–40 Mio reads/sample).
    Dispase, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dispase/product/Roche
    Average 86 stars, based on 1 article reviews
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    dispase - by Bioz Stars, 2021-03
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    Image Search Results


    PTPN13 or β-Catenin Silencing Increases Cell Adhesiveness and the Upregulation of Cell Membrane Adhesion Molecules (A) The proportion of Lin – (CD45 + Lin – ) and LSKs (CD45 + Lin – c-KIT + SCA1 + ) cells was analyzed in collagenase-/dispase-digested bones after BM had been flushed out (n = 4–5 different mice). The frequency of Lin – and LSKs strongly attached to the bone was higher in mice in which PTPN13 or β-catenin had been silenced. (B and C) PTPN13 or β-catenin was silenced in HEL cells. The levels of expression were analyzed by qRT-PCR (n = 5 or 8 independent experiments) (B) and immunoblotting (n = 3 independent experiments) (C). (D) Cell adhesiveness to collagen- or fibronectin-coated plates was analyzed in these cells (n = 5–7 independent experiments). (E) The expression of several genes coding for CAMs was analyzed by qRT-PCR (n = 4–9 independent experiments). (F) CDH1 promoter activity was analyzed by luciferase reporter assay in the presence or absence of a β-catenin stable mutant (β-catenin S33Y) (n = 4 independent experiments). Means ± SD are shown. See also Figure S4 .

    Journal: Stem Cell Reports

    Article Title: PTPN13 and β-Catenin Regulate the Quiescence of Hematopoietic Stem Cells and Their Interaction with the Bone Marrow Niche

    doi: 10.1016/j.stemcr.2015.08.003

    Figure Lengend Snippet: PTPN13 or β-Catenin Silencing Increases Cell Adhesiveness and the Upregulation of Cell Membrane Adhesion Molecules (A) The proportion of Lin – (CD45 + Lin – ) and LSKs (CD45 + Lin – c-KIT + SCA1 + ) cells was analyzed in collagenase-/dispase-digested bones after BM had been flushed out (n = 4–5 different mice). The frequency of Lin – and LSKs strongly attached to the bone was higher in mice in which PTPN13 or β-catenin had been silenced. (B and C) PTPN13 or β-catenin was silenced in HEL cells. The levels of expression were analyzed by qRT-PCR (n = 5 or 8 independent experiments) (B) and immunoblotting (n = 3 independent experiments) (C). (D) Cell adhesiveness to collagen- or fibronectin-coated plates was analyzed in these cells (n = 5–7 independent experiments). (E) The expression of several genes coding for CAMs was analyzed by qRT-PCR (n = 4–9 independent experiments). (F) CDH1 promoter activity was analyzed by luciferase reporter assay in the presence or absence of a β-catenin stable mutant (β-catenin S33Y) (n = 4 independent experiments). Means ± SD are shown. See also Figure S4 .

    Article Snippet: The bones were then digested with 0.02 mg/ml collagenase/dispase (Roche) for 2 hr at 37°C with constant agitation.

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Activity Assay, Luciferase, Reporter Assay, Mutagenesis

    Increased Adhesion of Mouse BM Progenitor Cells by PTPN13 or β-Catenin Silencing or by Treatment with Mesenchymal Cell-Conditioned Medium (A) Lin – progenitor cells were purified after flushing the BM of the transplanted animals (n = 4–5 different mice). (B) The hematopoietic cells that remained attached to the bone were obtained after collagenase/dispase digestion (n = 4–9 different mice). Both sources of cells were seeded in collagen- or fibronectin-coated plates, or in a monolayer of mouse mesenchymal stroma cells (mMSCs). 4 hr later, non-adherent cells were removed by gentle washing with PBS. The relative adhesion was monitored by comparing the percentage of GFP + adhered cells to the initial % GFP + cells. (C) Adhesion to collagen- or fibronectin-coated plates of Lin – cells treated with mesenchymal cell-conditioned medium (MC-CM) or with control medium during 4 hr (n = 3 independent experiments). (D) Messenger levels for PTPN13 and β-catenin in Lin – cells treated with MC-CM or control medium during 8 hr. (E) Messenger levels for several genes coding for CAMs treated with MC-CM or control medium during 8 hr (n = 5 independent experiments). Upregulated genes are shown in blue, and downregulated genes are in yellow. Means ± SD are shown.

    Journal: Stem Cell Reports

    Article Title: PTPN13 and β-Catenin Regulate the Quiescence of Hematopoietic Stem Cells and Their Interaction with the Bone Marrow Niche

    doi: 10.1016/j.stemcr.2015.08.003

    Figure Lengend Snippet: Increased Adhesion of Mouse BM Progenitor Cells by PTPN13 or β-Catenin Silencing or by Treatment with Mesenchymal Cell-Conditioned Medium (A) Lin – progenitor cells were purified after flushing the BM of the transplanted animals (n = 4–5 different mice). (B) The hematopoietic cells that remained attached to the bone were obtained after collagenase/dispase digestion (n = 4–9 different mice). Both sources of cells were seeded in collagen- or fibronectin-coated plates, or in a monolayer of mouse mesenchymal stroma cells (mMSCs). 4 hr later, non-adherent cells were removed by gentle washing with PBS. The relative adhesion was monitored by comparing the percentage of GFP + adhered cells to the initial % GFP + cells. (C) Adhesion to collagen- or fibronectin-coated plates of Lin – cells treated with mesenchymal cell-conditioned medium (MC-CM) or with control medium during 4 hr (n = 3 independent experiments). (D) Messenger levels for PTPN13 and β-catenin in Lin – cells treated with MC-CM or control medium during 8 hr. (E) Messenger levels for several genes coding for CAMs treated with MC-CM or control medium during 8 hr (n = 5 independent experiments). Upregulated genes are shown in blue, and downregulated genes are in yellow. Means ± SD are shown.

    Article Snippet: The bones were then digested with 0.02 mg/ml collagenase/dispase (Roche) for 2 hr at 37°C with constant agitation.

    Techniques: Purification, Mouse Assay

    Characterization of module thrombogenicity using whole-blood studies. ( a ) Clot formation times. The presence of HUVEC on the modules significantly increased the time to clot formation ( P = 1.4 × 10 −5 ) of slightly heparinized whole blood (0.75 units/ml) in a clotting test. In some cases, clot formation never actually occurred, and the test was terminated between 4,500 and 5,400 s; in these instances, the recorded time was the test termination time. Mean clot time is represented by the thick central line within the box. Open circles and stars represent outliers and extreme outliers, respectively. ( b ) Fresh whole blood (0.75 units/ml heparin) perfused through a HUVEC-covered modular construct (filled circles) maintains platelet levels no different from those measured in the absence of modules (open circles, flow circuit blank; includes polypropylene mesh required to keep modules in place). Blood perfusion through a control modular construct in which HUVEC have been removed by dispase–collagenase action (open squares), however, results in significant reductions in platelet number, indicating platelet activation and the thrombogenic response that occurs in the absence of HUVEC. Error bars indicate SEM ( n = 3, 4, and 7 for background, dispase-treated modular constructs, and HUVEC-covered modular constructs, respectively).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Vascularized organoid engineered by modular assembly enables blood perfusion

    doi: 10.1073/pnas.0602740103

    Figure Lengend Snippet: Characterization of module thrombogenicity using whole-blood studies. ( a ) Clot formation times. The presence of HUVEC on the modules significantly increased the time to clot formation ( P = 1.4 × 10 −5 ) of slightly heparinized whole blood (0.75 units/ml) in a clotting test. In some cases, clot formation never actually occurred, and the test was terminated between 4,500 and 5,400 s; in these instances, the recorded time was the test termination time. Mean clot time is represented by the thick central line within the box. Open circles and stars represent outliers and extreme outliers, respectively. ( b ) Fresh whole blood (0.75 units/ml heparin) perfused through a HUVEC-covered modular construct (filled circles) maintains platelet levels no different from those measured in the absence of modules (open circles, flow circuit blank; includes polypropylene mesh required to keep modules in place). Blood perfusion through a control modular construct in which HUVEC have been removed by dispase–collagenase action (open squares), however, results in significant reductions in platelet number, indicating platelet activation and the thrombogenic response that occurs in the absence of HUVEC. Error bars indicate SEM ( n = 3, 4, and 7 for background, dispase-treated modular constructs, and HUVEC-covered modular constructs, respectively).

    Article Snippet: Constructs were assembled from HUVEC-covered modules that were untreated or treated for 15 min in 100 mg/ml collagenase dispase solution (Roche, Mississauga, ON, Canada) to remove all HUVEC from the surface (for control modules), yet retain the size and stiffness of the contracted collagen.

    Techniques: Coagulation, Construct, Flow Cytometry, Activation Assay

    Overview of retinal endothelial cell isolation for RNA sequencing P5 retinas from wild‐type mice were digested with collagenase and dispase. CD31 and CD34 double‐positive endothelial cells were isolated by FACS, pooled into two samples, and analyzed by RNA‐Seq on an Illumina HiSeq 4000 sequencer generating 50 bp single‐end reads ( ca. 30–40 Mio reads/sample).

    Journal: EMBO Reports

    Article Title: EVL regulates VEGF receptor‐2 internalization and signaling in developmental angiogenesis

    doi: 10.15252/embr.201948961

    Figure Lengend Snippet: Overview of retinal endothelial cell isolation for RNA sequencing P5 retinas from wild‐type mice were digested with collagenase and dispase. CD31 and CD34 double‐positive endothelial cells were isolated by FACS, pooled into two samples, and analyzed by RNA‐Seq on an Illumina HiSeq 4000 sequencer generating 50 bp single‐end reads ( ca. 30–40 Mio reads/sample).

    Article Snippet: The remaining pellet was further digested with Collagenase, Dispase (Roche), and DNase I (Worthington) in buffer A for 15 min at 37°C.

    Techniques: Cell Isolation, RNA Sequencing Assay, Mouse Assay, Isolation, FACS