p14 pancreas required collagenase digestion  (Millipore)

 
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    Name:
    Collagenase D
    Description:
    Collagenase D is prepared from Clostridium histolyticum cultures by filtration ammonium sulfate precipitation dialysis and lyophilization
    Catalog Number:
    COLLD-RO
    Price:
    None
    Applications:
    Collagenase from C. histolyticum is widely used for the disaggregation of many types of tissues (e.g., lung, heart, muscle, bone, adipose tissue, liver, kidney, cartilage, mammary gland, placentae, blood vessels, brain, tumors) and for the preparation of single cell suspensions for the establishment of primary cell culture systems. Collagenase D is recommended when functionality and integrity of cell-surface proteins are important.Clostridium collagenase from Roche has been used to prepare cells from many types of tissue, such as hepatocytes, adipocytes, pancreatic islets, epithelial cells, muscle cells, endothelial cells, etc. However, suitability of each lot of the enzyme for disruption of a particular tissue should be determined empirically.
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    Structured Review

    Millipore p14 pancreas required collagenase digestion
    Collagenase D
    Collagenase D is prepared from Clostridium histolyticum cultures by filtration ammonium sulfate precipitation dialysis and lyophilization
    https://www.bioz.com/result/p14 pancreas required collagenase digestion/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p14 pancreas required collagenase digestion - by Bioz Stars, 2021-07
    97/100 stars

    Images

    1) Product Images from "Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability"

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    doi: 10.1152/ajpendo.00260.2017

    Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.
    Figure Legend Snippet: Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.

    Techniques Used: Expressing, Functional Assay

    Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.
    Figure Legend Snippet: Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.

    Techniques Used:

    Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.
    Figure Legend Snippet: Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.

    Techniques Used: Expressing, Functional Assay

    Related Articles

    Isolation:

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability
    Article Snippet: .. Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient. ..

    Incubation:

    Article Title: Polarization of macrophages in the tumor microenvironment is influenced by EGFR signaling within colon cancer cells
    Article Snippet: Flow cytometry Isolation of primary mouse macrophages from colon was performed as described previously [ ]. .. Briefly, colon tissues were cut into small pieces (1-2 mm) and incubated in 10 mL PRMI 1640 medium with 10 mM HEPES and 5% fetal bovine serum containing 10 mg (1 mg/mL) collagenase D (Sigma-Aldrich), 10 mg (1 mg/mL) dispase II (Roche, Germany), and 100 μL 10 mg/ml DNase I (100 μg/mL) (Sigma-Aldrich) for 30-45 min in a shaking incubator at 37°C. .. For Ana-1 and BMDM cells, single-cell suspensions from mouse colon tissues were incubated with APC-anti-mouse CD11b and FITC-anti-mouse F4/80 (Affymetrix eBioscience, USA), or APC-anti-mouse F4/80 and FITC-anti-mouse CD206 (MMR) (Biolegend, San Diego, CA) antibodies.

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

    Flow Cytometry:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Cytometry:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Lysis:

    Article Title: Dysregulated Cytokine Expression by CD4+ T cells from Post-Septic Mice Modulates both Th1 and Th2-Mediated Granulomatous Lung Inflammation
    Article Snippet: .. Flow cytometry Collagenase-digested (Sigma-Aldrich, St. Louis, MO) lung lobes and undigested lymph nodes from animals were processed into single cell suspensions by processing tissues through sterile 40-mm filters, and ammonium chloride lysis buffer was used to eliminate erythrocytes. .. Cells were stained with the following fluorescent antibodies in flow cytometry buffer (phosphate buffered saline, 1% w/v bovine serum albumin, 0.05% w/v sodium azide): Purified αCD16/32 (Fc Block) (2.4G2, BD Biosciences, San Jose, CA), FITC-αCD8a (53–6.7, BD Biosciences), FITC-αCD3ε (145-2C11, BD Biosciences), PeCy7-αCD45 (Ly5, eBioscience, San Diego, CA), PeCy7-αCD3ε (17A2, BioLegend, San Diego, CA), Pacific Blue- (RM4-5, BioLegend) or Pacific Orange (RM4-5, Invitrogen, Carlsbad, CA) -αCD4, and Pacific Blue-αCD45 (30-F11, BioLegend).

    Mouse Assay:

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

    Infection:

    Article Title: Resolution of a chronic viral infection after interleukin-10 receptor blockade
    Article Snippet: .. Splenocytes from mice infected 7 d earlier with LCMV Armstrong or LCMV clone 13 with or without anti–IL-10R antibody treatment were incubated with HBSS medium containing 0.5 mg/ml collagenase D (Sigma-Aldrich) at 37°C, 5% CO2 for 30 min. 0.01 M EDTA was added to disrupt T cell–DC complexes. .. Next, cells were depleted of CD3-expressing cells (Dynal CD3-beads; Dynal), incubated with CD11c microbeads (Miltenyi Biotec), and positively selected using MACS columns.

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  • About
  • News
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  • 97
    Millipore p14 pancreas required collagenase digestion
    Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in <t>P14</t> islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.
    P14 Pancreas Required Collagenase Digestion, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p14 pancreas required collagenase digestion/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    p14 pancreas required collagenase digestion - by Bioz Stars, 2021-07
    97/100 stars
      Buy from Supplier

    Image Search Results


    Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Double heterozygotes have significant alterations in postnatal islet gene expression. A : expression of select endocrine development and functional genes in P14 islets represented in a heatmap. Green, increased gene expression; red, reduced gene expression. Blue boxes, controls; green boxes, PH; red boxes, OH; black boxes, DH. B – D : relative expression levels of hormones ( B ), regulatory transcription factors ( C ), and islet functional genes ( D ) ( n = 4). One symbol, P ≤ 0.05; two symbols, P ≤ 0.01; three symbols, P ≤ 0.001. +Control (Con) vs. PH; #DH vs. PH; ^OH vs. DH; *Con vs. DH by 2-way ANOVA with Tukey’s correction for multiple comparisons or Kruskal-Wallis with Dunn’s correction for multiple comparisons.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques: Expressing, Functional Assay

    Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Islets of double heterozygotes show altered postnatal maturation. A : P14 DH islets lack a distinct β-cell core and α-cell mantle (green, insulin; red, glucagon; blue, DAPI). B – D : quantification of the percentage of islets with a mixed phenotype ( B ) and the percentage of nonmantle α-cells in each genotype at P14 ( C ), without a change in β- to α-cell ratio ( D ) ( n = 5–7). E and F : urocortin3 mRNA ( E ) and protein ( F ) are reduced in PH and DH islets ( n = 3 for mRNA, images representative of 6 animals). Scale bar: 100 μm. * P ≤ 0.05.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques:

    Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: Cooperative function of Pdx1 and Oc1 in multipotent pancreatic progenitors impacts postnatal islet maturation and adaptability

    doi: 10.1152/ajpendo.00260.2017

    Figure Lengend Snippet: Double heterozygosity negatively impacts expression of regulatory and functional factors in postnatal islets. A : Nkx6.1 (red) is detected in fewer β-cells (insulin, green) and has reduced intensity in P14 DH. Arrowheads in DH indicate Nkx6.1 + cells. B : quantification of A ( n = 4–6). C : Glut2 expression (red) is weaker and more diffuse (E-cadherin, green) in β-cells (insulin, yellow) of DH. DAPI is in blue ( n = 4–6). Scale bar: 100 μm. **** P ≤ 0.0001 by 1-Way ANOVA with Tukey’s correction for multiple comparisons.

    Article Snippet: Isolation of islets from P14 pancreas required collagenase digestion of whole dissected pancreas at 37°C and hand-picking of islets away from exocrine tissue following a Histopaque-1077 (Sigma-Aldrich) gradient.

    Techniques: Expressing, Functional Assay