Avidin-Biotin Assay:Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence Chemically Induced Lung Tumor Models A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee. .. For single agent carcinogenesis, male A/J, FVB, and B6 mice were injected i.p. (intraperitoneally) with 1 mg urethane (ethyl carbamate; Sigma, St. Louis, MO)/g body wt dissolved in sterile 0.9% NaCl as previously described 31 ; control mice received a single saline i.p. injection.
Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence .. A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee.
Activation Assay:Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence Chemically Induced Lung Tumor Models A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee. .. For single agent carcinogenesis, male A/J, FVB, and B6 mice were injected i.p. (intraperitoneally) with 1 mg urethane (ethyl carbamate; Sigma, St. Louis, MO)/g body wt dissolved in sterile 0.9% NaCl as previously described 31 ; control mice received a single saline i.p. injection.
Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence .. A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee.
Immunofluorescence:Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence Chemically Induced Lung Tumor Models A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee. .. For single agent carcinogenesis, male A/J, FVB, and B6 mice were injected i.p. (intraperitoneally) with 1 mg urethane (ethyl carbamate; Sigma, St. Louis, MO)/g body wt dissolved in sterile 0.9% NaCl as previously described 31 ; control mice received a single saline i.p. injection.
Article Title: Tumor Progression Stage and Anatomical Site Regulate Tumor-Associated Macrophage and Bone Marrow-Derived Monocyte Polarization
Article Snippet: .. Samples were first blocked with avidin-biotin solution (Vector Laboratories, Burlingame, CA), followed by exposure to the primary and biotinylated secondary antibodies and detection with fluorescent avidin conjugates. table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Dilution Secondary Sample type Fluorophore/color F4/80–polycolonal (Caltag Laboratories, Carlsbad, CA) 1:100 Rat BAL, human and mouse pulmonary macrophages, peritoneal macrophages Fluorescein/green CD-68–monocolonal (Dako, Glostrup, Denmark) 1:100 Mouse BDMCs, xenograft TAMs, prostate TAMs Fluorescein/green Arginase I–polyclonal (Santa Cruz Biotech, Santa Cruz, CA) 1:100 Goat Murine M2 activation AMCA/blue CD-206/Mannose receptor–monoclonal (Serotec, Oxford, UK) 1:100 Human Human M2 activation AMCA/blue iNOS–polyclonal (BD Pharmagen, San Diego, CA) 1:100 Rabbit Mouse and human M1 activation Rhodamine/red Open in a separate window Antibodies and Conditions for Immunofluorescence .. A/J, FVB, BALB/cByJ (hereafter, cBy), BALB/cJ, and C57BL/6J (hereafter, B6) male and female mice 4 to 6 weeks of age were purchased from Jackson Laboratory (Bar Harbor, ME), housed on hardwood bedding with 12-hour light/dark cycles, and fed standard rodent diet (Harlan Teklad, Madison, WI) at the Center for Laboratory Animal Care at the University of Colorado Denver under a protocol approved by the University of Colorado Institutional Animal Care and Use Committee.
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