collagenase d  (Roche)


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    Structured Review

    Roche collagenase d
    Collagenase D, 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 d/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase d - by Bioz Stars, 2021-04
    86/100 stars

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    other:

    Article Title: Fetal-derived macrophages dominate in adult mammary glands
    Article Snippet: MG was digested with 1 mg/ml collagenase D (Roche, 1108886601) and 50 µg/ml DNase 1 (Roche, 10104159001) for 60 min at +37 °C.

    Article Title: Dental cell type atlas reveals stem and differentiated cell types in mouse and human teeth
    Article Snippet: The pulp was then resuspended in 5 ml of Collagenase D (0.5U/ml, Roche, 11088866001) and Dispase II (1.5U/ml, Roche, 4942078001).

    Article Title: Absence of MHC-II expression by lymph node stromal cells results in autoimmunity
    Article Snippet: The remaining tissue was digested twice in RPMI containing 1 mg/ml of collagenase D (Roche) for 30 min at 37°C in a shaking incubator.

    Cell Culture:

    Article Title: RSU-1 interaction with prohibitin-2 links cell–extracellular matrix detachment to downregulation of ERK signaling
    Article Snippet: Cells (as specified in each experiment) were harvested with trypsin, washed, and mixed with collagen I (the final cell density = 5.0 × 105 cells/ml and the final concentration of collagen I = 1.0 mg/ml), and pH was adjusted to 7.4 with 1 M NaOH. .. The cells were cultured in 3D collagen I gels for 24 h and then transferred into tubes and treated with 1 mg/ml collagenase D (Roche, 37334226) for 0.5 h at 37 °C. .. The cells were collected by centrifugation at 300g for 3 min and lysed with the radio-immunoprecipitation assay buffer for further analyses.

    Incubation:

    Article Title: A Circular RNA Binds To and Activates AKT Phosphorylation and Nuclear Localization Reducing Apoptosis and Enhancing Cardiac Repair
    Article Snippet: .. The tissue fragments were transferred into a new tube and incubated in 25 ml Tyrode solution containing 0.012 g Collagenase D (Roche Diagnostics, 1-088-882), 0.009 g Collagenase B, and 0.001 g Protease XIV at 37°C for 30 min. .. Cell pellet was re-suspend in DMEM/F12 medium containing 10% FBS and 20 mM BDM and plated onto10-cm cell culture dishes and incubated for 2 h. This pre-plating step removed fibroblasts and endothelial cells, which adhered to the uncoated dish.

    Article Title: The Combined Use of Melatonin and an Indoleamine 2,3-Dioxygenase-1 Inhibitor Enhances Vaccine-Induced Protective Cellular Immunity to HPV16-Associated Tumors
    Article Snippet: Subsequently, cells were washed and distributed in 96-well U-bottom plates for further staining. .. Tumor masses were minced with scissors and submitted to enzymatic digestion with 0.22 u/mL of collagenase D (#11088866001, Roche Diagnostics) at 37°C for 1 h, stirring gently every 10 min. After the incubation period, the enzyme was inactivated with 5 mM EDTA at room temperature for 5 min. Then, the samples gentle resuspended in R10 and filtered on a 70 μm cell strainer (Easy strainer Greiner Bio One). .. After centrifugation, the pelleted cells were resuspended in R10 and filtered on a 40 μm cell strainer.

    Article Title: The neonatal Fc receptor is a pan-echovirus receptor
    Article Snippet: For collection of primary cells, mice were killed according to institution standards and ears and tail were removed, incubated in 70% ethanol for 5 min, and then rinsed twice in PBS + 50 μg/mL kanamycin for 5 min. .. Hair was removed and tissue was cut into small pieces and incubated in 9.4 mg/mL collagenase D (11088858001, Roche) and 1.2 mg/mL pronase (1088858001, Roche) in complete DMEM at 37 °C with shaking at 200 rpm for 90 min. .. The resulting cell suspensions were filtered through 70-μM cell strainers, collected at 580 g, resuspended in complete DMEM containing 10 units penicillin and 10 μg streptomycin/mL and 250 ng/mL amphotericin B and cultured at 37 °C in a humidified 5% CO2 incubator.

    Mouse Assay:

    Article Title: The Nuclear Orphan Receptor NR2F6 Is a Central Checkpoint for Cancer Immune Surveillance
    Article Snippet: Preparation of Tumor-Infiltrating Cells Mononuclear infiltrating cells were isolated from both subcutaneous and autochthonous tumors at the indicated time points. .. Briefly, tumor tissues from sacrificed mice were prepared by mechanical disruption followed by digestion for 45 min with collagenase D (2.5 mg/ml; Roche, 11088858001) and DNase I (1 mg/ml; Roche, 11284932001) at 37°C. .. Digested tissues were incubated 5 min at 37°C with EDTA (0.5 M) to prevent DC/T cell aggregates and mashed through filters.

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    Roche collagenase d
    Absence of donor APC-derived IL-23 significantly attenuates proinflammatory cytokine production in the colon . (A,B) Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 BM (10 × 10 6 ) and spleen cells (0.4-0.6 × 10 6 ) (■) or IL-23 −/−  BM (10 × 10 6 ) and spleen cells (□) adjusted to yield an equivalent number of mature T cells. Cohorts of mice (7-9/group) were killed at the indicated time points (days 4, 7, 14, 21, and 28), and segments of colon tissue from individual mice were cultured overnight in media. Colonic tissue supernatants were collected and analyzed for the amount of IL-23p19 (A) and IL-12p70 (B) by ELISA and multiplex, respectively. Data are presented as mean amount of cytokine divided by the weight of cultured colon tissue (± SEM) and are cumulative results from 2 experiments. (C) Groups of mice were transplanted as in panels A and B and killed 28 days posttransplantation. Colons (n = 4-5/group per experiment) were pooled and digested with collagenase D. Total number of isolated LPMCs that were CD11c +  CD11b +  or CD11b +  CD11c −  in Balb/c recipients of B6 BM and spleen cells (■) or IL-23 −/−  BM and spleen cells (□) is depicted. Data are derived from 3 independent experiments and are presented as the mean cell number (×1000) per mouse (± SEM) (D). Lethally irradiated Balb/c mice were transplanted with B6 BM plus spleen cells (n = 10, ■) or IL-23 −/−  BM and spleen cells adjusted to yield equivalent numbers of T cells (n = 9, □). Mice were killed 21-29 days posttransplantation, and colon explant tissue was assayed for levels of proinflammatory cytokines by multiplex. Data are derived from cumulative results from 2 experiments and are presented as the mean amount of cytokine divided by the weight of cultured colon tissue (± SEM). Statistics: * P  ≤ .05; ** P
    Collagenase D, 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 d/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase d - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

    86
    Roche collagenase d solution
    Nucleated cell number, harvested cell number, and cell morphology at passage 0. (A) The numbers of nucleated cells tissue weight and per bone marrow volume. Synovium, infrapatellar fat pad, and adipose tissues were digested with <t>collagenase</t> D solution, and nucleated cell numbers were counted for these three tissues as well as for bone marrow. (B) Harvested cell number after 14 days of culture. The nucleated cells were plated at the determined optimal plating density and after 14 days of culture, the numbers of harvested cells were counted. Values are the mean ± standard deviation (n = 6). Statistical analyses were carried out only for the three solid tissues. **, p
    Collagenase D 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 d solution/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    collagenase d solution - by Bioz Stars, 2021-04
    86/100 stars
      Buy from Supplier

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    Absence of donor APC-derived IL-23 significantly attenuates proinflammatory cytokine production in the colon . (A,B) Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 BM (10 × 10 6 ) and spleen cells (0.4-0.6 × 10 6 ) (■) or IL-23 −/−  BM (10 × 10 6 ) and spleen cells (□) adjusted to yield an equivalent number of mature T cells. Cohorts of mice (7-9/group) were killed at the indicated time points (days 4, 7, 14, 21, and 28), and segments of colon tissue from individual mice were cultured overnight in media. Colonic tissue supernatants were collected and analyzed for the amount of IL-23p19 (A) and IL-12p70 (B) by ELISA and multiplex, respectively. Data are presented as mean amount of cytokine divided by the weight of cultured colon tissue (± SEM) and are cumulative results from 2 experiments. (C) Groups of mice were transplanted as in panels A and B and killed 28 days posttransplantation. Colons (n = 4-5/group per experiment) were pooled and digested with collagenase D. Total number of isolated LPMCs that were CD11c +  CD11b +  or CD11b +  CD11c −  in Balb/c recipients of B6 BM and spleen cells (■) or IL-23 −/−  BM and spleen cells (□) is depicted. Data are derived from 3 independent experiments and are presented as the mean cell number (×1000) per mouse (± SEM) (D). Lethally irradiated Balb/c mice were transplanted with B6 BM plus spleen cells (n = 10, ■) or IL-23 −/−  BM and spleen cells adjusted to yield equivalent numbers of T cells (n = 9, □). Mice were killed 21-29 days posttransplantation, and colon explant tissue was assayed for levels of proinflammatory cytokines by multiplex. Data are derived from cumulative results from 2 experiments and are presented as the mean amount of cytokine divided by the weight of cultured colon tissue (± SEM). Statistics: * P  ≤ .05; ** P

    Journal: Blood

    Article Title: Interleukin-23 secretion by donor antigen-presenting cells is critical for organ-specific pathology in graft-versus-host disease

    doi: 10.1182/blood-2008-08-175448

    Figure Lengend Snippet: Absence of donor APC-derived IL-23 significantly attenuates proinflammatory cytokine production in the colon . (A,B) Lethally irradiated (900 cGy) Balb/c mice were transplanted with B6 BM (10 × 10 6 ) and spleen cells (0.4-0.6 × 10 6 ) (■) or IL-23 −/− BM (10 × 10 6 ) and spleen cells (□) adjusted to yield an equivalent number of mature T cells. Cohorts of mice (7-9/group) were killed at the indicated time points (days 4, 7, 14, 21, and 28), and segments of colon tissue from individual mice were cultured overnight in media. Colonic tissue supernatants were collected and analyzed for the amount of IL-23p19 (A) and IL-12p70 (B) by ELISA and multiplex, respectively. Data are presented as mean amount of cytokine divided by the weight of cultured colon tissue (± SEM) and are cumulative results from 2 experiments. (C) Groups of mice were transplanted as in panels A and B and killed 28 days posttransplantation. Colons (n = 4-5/group per experiment) were pooled and digested with collagenase D. Total number of isolated LPMCs that were CD11c + CD11b + or CD11b + CD11c − in Balb/c recipients of B6 BM and spleen cells (■) or IL-23 −/− BM and spleen cells (□) is depicted. Data are derived from 3 independent experiments and are presented as the mean cell number (×1000) per mouse (± SEM) (D). Lethally irradiated Balb/c mice were transplanted with B6 BM plus spleen cells (n = 10, ■) or IL-23 −/− BM and spleen cells adjusted to yield equivalent numbers of T cells (n = 9, □). Mice were killed 21-29 days posttransplantation, and colon explant tissue was assayed for levels of proinflammatory cytokines by multiplex. Data are derived from cumulative results from 2 experiments and are presented as the mean amount of cytokine divided by the weight of cultured colon tissue (± SEM). Statistics: * P ≤ .05; ** P

    Article Snippet: To isolate lamina propria mononuclear cells (LPMCs), pooled colons were incubated in Hank balanced salt solution (HBSS) buffer (Gibco-BRL Life Technologies) supplemented with 5% fetal bovine serum (FBS), 0.05 mM ethylenediaminetetraacetic acid (EDTA), 0.6 mM N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (HEPES) buffer, 100 U/mL penicillin/100 μg/mL streptomycin (Gibco-BRL Life Technologies), and 15 μg/mL dithiothreitol (Invitrogen, Carlsbad, CA) at 37°C for 30 minutes and subsequently digested in a solution of 0.2 mg/mL collagenase D (Roche Diagnostics, Indianapolis, IN) in RPMI medium for 1 hour at 37°C.

    Techniques: Derivative Assay, Irradiation, Mouse Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Multiplex Assay, Isolation

    I.t. SLR14 delivery enhances tumor infiltration of cytotoxic T lymphocytes and myeloid cells. Subcutaneous YMR1.7 melanoma was established in C57BL/6J mice and i.t. treated with vehicle, SLR14, or no treatment. 3 d after the fifth treatment, tumors were harvested and digested with 0.5 mg/ml Collagenase D and 40 µg/ml DNase I. Single-cell suspensions were prepared for flow cytometry analysis. (A) Percentages (top) and quantities (bottom) of tumor-infiltrating CD45 + , CD11b + , CD8 + , CD4 + , FoxP3 + CD4 + , or NK1.1 + cells in each group. All T cells were CD44 + . The cell numbers were normalized based on the tumor weight. Error bars = SD. 1, no treatment; 2, vehicle; 3, SLR14. (B) The ratio of tumor-infiltrating CD8 + T cells/CD4 + T cells or CD8 + T cells/CD4 + FoxP3 + T reg cells in each group. Error bars = SD. (C) Subcutaneous YMR1.7 melanoma growth in RAG1 −/− mice treated with vehicle or SLR14. Treatment protocol was the same as described in Fig. 1 A . Left: Tumor growth curves (error bars = SD) for each group of mice. Right: Tumor growth curves of individual mice in each group. (D and E) IFNγ, TNFα, and GzmB productions of tumor-infiltrating CD8 + T lymphocytes after i.t. treatment (error bars = SD). Five mice per group. Unpaired t test was used for statistical analysis. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Intratumoral delivery of RIG-I agonist SLR14 induces robust antitumor responses

    doi: 10.1084/jem.20190801

    Figure Lengend Snippet: I.t. SLR14 delivery enhances tumor infiltration of cytotoxic T lymphocytes and myeloid cells. Subcutaneous YMR1.7 melanoma was established in C57BL/6J mice and i.t. treated with vehicle, SLR14, or no treatment. 3 d after the fifth treatment, tumors were harvested and digested with 0.5 mg/ml Collagenase D and 40 µg/ml DNase I. Single-cell suspensions were prepared for flow cytometry analysis. (A) Percentages (top) and quantities (bottom) of tumor-infiltrating CD45 + , CD11b + , CD8 + , CD4 + , FoxP3 + CD4 + , or NK1.1 + cells in each group. All T cells were CD44 + . The cell numbers were normalized based on the tumor weight. Error bars = SD. 1, no treatment; 2, vehicle; 3, SLR14. (B) The ratio of tumor-infiltrating CD8 + T cells/CD4 + T cells or CD8 + T cells/CD4 + FoxP3 + T reg cells in each group. Error bars = SD. (C) Subcutaneous YMR1.7 melanoma growth in RAG1 −/− mice treated with vehicle or SLR14. Treatment protocol was the same as described in Fig. 1 A . Left: Tumor growth curves (error bars = SD) for each group of mice. Right: Tumor growth curves of individual mice in each group. (D and E) IFNγ, TNFα, and GzmB productions of tumor-infiltrating CD8 + T lymphocytes after i.t. treatment (error bars = SD). Five mice per group. Unpaired t test was used for statistical analysis. *, P

    Article Snippet: Tumor digestion and flow cytometry analysis Tumors were harvested, cut into small pieces with surgical scissors and sharp blade, and then digested in HBSS containing 0.5 mg/ml Collagenase D (Roche) and 40 µg/ml DNase I (Roche) in a 37°C shaker for 20–30 min. Digestion was stopped by adding 0.5 mg/ml EDTA in HBSS, and single-cell suspensions were prepared for antibody staining.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    Impaired activation of PI3Kδ−/− CTLs by tumor cells. A. 1×10 6 MC38 colon adenocarcinoma cells were injected subcutaneously into the flanks of WT and PI3Kδ−/− animals. Fifteen days later tumor weights were analyzed. Depicted is the summary of 3 independent experiments (WT: 0.19±0.19 g; versus PI3Kδ−/− : 1.16±0.33 g, n = 12, values represent mean±SD). B. Representative pictures of MC38 solid tumors grown in WT and PI3Kδ−/− animals. C. After removal, tumors were minced and digested with collagenase D and DNase I. Tumor-infiltrating CD3+CD8+ CTLs were quantified using flow cytometry (WT: 1.9±0.5%; versus PI3Kδ−/− : 1.8±0.7%, n = 8, values represent mean±SD). D., E. Representative histograms showing expression levels of CD44, CD45RB, CD62L and CD69 on splenic CD3+CD8+ T cells from untreated WT and PI3Kδ−/− mice (D) and on MC38 tumor-infiltrating CD3+CD8+ CTLs of WT and PI3Kδ−/− mice (E). F. Summary of the expression levels of CD44 (WT: median = 19040, IQR = 16670–24220; versus PI3Kδ−/− : median = 15053, IQR = 12750–26440, n = 8), CD45RB (WT: median = 3222, IQR = 2907–4142; versus PI3Kδ−/− : median = 3945, IQR = 2579–5833, n = 8), CD62L (WT: median = 194, IQR = 146–219; versus PI3Kδ−/− : median = 228, IQR = 187–333, n = 8, Mann-Whitney test: p = 0.0572 ) and CD69 (WT: median = 2673, IQR = 1825–7606; versus PI3Kδ−/− : median = 1428, IQR = 1334–1698, n = 8, Mann-Whitney test: p = 0.0037 ) on tumor-infiltrating WT and PI3Kδ−/− CD3+CD8+ CTLs.

    Journal: PLoS ONE

    Article Title: PI3K? Is Essential for Tumor Clearance Mediated by Cytotoxic T Lymphocytes

    doi: 10.1371/journal.pone.0040852

    Figure Lengend Snippet: Impaired activation of PI3Kδ−/− CTLs by tumor cells. A. 1×10 6 MC38 colon adenocarcinoma cells were injected subcutaneously into the flanks of WT and PI3Kδ−/− animals. Fifteen days later tumor weights were analyzed. Depicted is the summary of 3 independent experiments (WT: 0.19±0.19 g; versus PI3Kδ−/− : 1.16±0.33 g, n = 12, values represent mean±SD). B. Representative pictures of MC38 solid tumors grown in WT and PI3Kδ−/− animals. C. After removal, tumors were minced and digested with collagenase D and DNase I. Tumor-infiltrating CD3+CD8+ CTLs were quantified using flow cytometry (WT: 1.9±0.5%; versus PI3Kδ−/− : 1.8±0.7%, n = 8, values represent mean±SD). D., E. Representative histograms showing expression levels of CD44, CD45RB, CD62L and CD69 on splenic CD3+CD8+ T cells from untreated WT and PI3Kδ−/− mice (D) and on MC38 tumor-infiltrating CD3+CD8+ CTLs of WT and PI3Kδ−/− mice (E). F. Summary of the expression levels of CD44 (WT: median = 19040, IQR = 16670–24220; versus PI3Kδ−/− : median = 15053, IQR = 12750–26440, n = 8), CD45RB (WT: median = 3222, IQR = 2907–4142; versus PI3Kδ−/− : median = 3945, IQR = 2579–5833, n = 8), CD62L (WT: median = 194, IQR = 146–219; versus PI3Kδ−/− : median = 228, IQR = 187–333, n = 8, Mann-Whitney test: p = 0.0572 ) and CD69 (WT: median = 2673, IQR = 1825–7606; versus PI3Kδ−/− : median = 1428, IQR = 1334–1698, n = 8, Mann-Whitney test: p = 0.0037 ) on tumor-infiltrating WT and PI3Kδ−/− CD3+CD8+ CTLs.

    Article Snippet: For flow cytometric analysis the tumors were minced and digested with 1 mg/ml collagenase D (Roche Applied Sciences) and 0.05 mg/ml DNase I (Roche Applied Sciences) for 1h at 37°C.

    Techniques: Activation Assay, Injection, Flow Cytometry, Cytometry, Expressing, Mouse Assay, MANN-WHITNEY

    Nucleated cell number, harvested cell number, and cell morphology at passage 0. (A) The numbers of nucleated cells tissue weight and per bone marrow volume. Synovium, infrapatellar fat pad, and adipose tissues were digested with collagenase D solution, and nucleated cell numbers were counted for these three tissues as well as for bone marrow. (B) Harvested cell number after 14 days of culture. The nucleated cells were plated at the determined optimal plating density and after 14 days of culture, the numbers of harvested cells were counted. Values are the mean ± standard deviation (n = 6). Statistical analyses were carried out only for the three solid tissues. **, p

    Journal: PLoS ONE

    Article Title: Canine mesenchymal stem cells from synovium have a higher chondrogenic potential than those from infrapatellar fat pad, adipose tissue, and bone marrow

    doi: 10.1371/journal.pone.0202922

    Figure Lengend Snippet: Nucleated cell number, harvested cell number, and cell morphology at passage 0. (A) The numbers of nucleated cells tissue weight and per bone marrow volume. Synovium, infrapatellar fat pad, and adipose tissues were digested with collagenase D solution, and nucleated cell numbers were counted for these three tissues as well as for bone marrow. (B) Harvested cell number after 14 days of culture. The nucleated cells were plated at the determined optimal plating density and after 14 days of culture, the numbers of harvested cells were counted. Values are the mean ± standard deviation (n = 6). Statistical analyses were carried out only for the three solid tissues. **, p

    Article Snippet: Synovium, infrapatellar fat pad, and adipose tissues were minced and digested at 37°C for 3 hours in a 3 mg/ml collagenase D solution (Roche Diagnostics, Mannheim, Germany) in α-minimal essential medium (αMEM; Invitrogen, Carlsbad, CA, USA).

    Techniques: Standard Deviation