cd45 percp rat igg2b  (R&D Systems)

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Altered T-Cell Balance in Lymphoid Organs of a Mouse Model of Colorectal Cancer

doi: 10.1369/0022155416672418

Figure Lengend Snippet: Antibody Information.

Article Snippet: After 72 hr, supernatants were collected and cells were rested for 24 hr with IL-2 (10 ng/ml) before staining as described below. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antigen Clone Conjugate Raised In Isotype Supplier Catalog Number Dilution/Concentration Purpose CD3e 145-2C11 NA Armenian hamster IgG1, κ BD Biosciences 553058 1:500 Cell stimulation CD28 37.51 NA Syrian hamster IgG2, λ 1 BD Biosciences 553294 1:2000 Cell stimulation CD45 30-F11 PerCP-Cy5.5 Rat IgG2b, κ BD Biosciences 561869 1:400 Flow cytometry CD4 RM4-5 Allophycocyanin Rat IgG1a, κ BD Biosciences 553051 1:100 Flow cytometry CD8a 53-6.7 Allophycocyanin Rat IgG1a, κ BD Biosciences 553035 1:100 Flow cytometry IFN-γ XMG1.1 FITC Rat IgG1, κ BD Biosciences 554411 1:100 Flow cytometry IL-17 TC11-18H10 PE Rat IgG1, κ BD Biosciences 559502 1:200 Flow cytometry IL-4 11B11 NA Rat IgG1, κ BioLegend 504107 10 µg/ml Cell polarization IFN-γ XMG1.2 NA Rat IgG1, κ BioLegend 505811 10 µg/ml Cell polarization IFN-γ XMG1.2 Biotin Rat IgG1, κ BioLegend 505803 1 µg/ml ELISA, 1:100 IF IF and ELISA IFN-γ R4-6A2 NA Rat IgG1, κ BioLegend 505704 0.5 µg/ml ELISA IL-17A TC11-18H10.1 NA Rat IgG1, κ BioLegend 506901 1 µg/ml ELISA IL-17A TC11-8H4 Biotin Rat IgG1, κ BioLegend 507001 0.5 µg/ml ELISA Granzyme B NGZB PE Rat IgG1a, κ eBioscience 12-8898 1:100 Flow cytometry Rat Ig Polyclonal Biotin Donkey Polyclonal Jackson ImmunoResearch 112-065-003 1:400 IF Streptavidin NA Horseradish peroxidase NA NA Jackson ImmunoResearch 016-030-084 1:1000 ELISA Streptavidin NA Alexa Fluor 488 NA NA Thermo Fisher Scientific (Carlsbad, CA) S-32354 1:400 IF CD8a 53-6.7 NA Rat IgG1a, χ Santa Cruz sc-18913 1:500 IF Open in a separate window Abbreviations: IFN-γ, interferon gamma; IL, interleukin; PE, phycoerythrin; IF, immunofluorescence; NA, not applicable.

Techniques: Cell Stimulation, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Histochemistry and Cytochemistry

Article Title: Altered T-Cell Balance in Lymphoid Organs of a Mouse Model of Colorectal Cancer

doi: 10.1369/0022155416672418

Figure Lengend Snippet: Decreased levels of IFN-γ+IL-17+ double-positive CD4+ T cells in the intestines of ApcMin/+ mice. (A) Percentages of total CD3+CD4+ T cells, IFN-γ+ single-positive, IL-17+ single-positive, and IFN-γ+IL-17+ double-positive CD4+ T cells from spleen, MLN, PPs, SILP, CoLP, and small intestinal IEL of Apc+/+ and ApcMin/+ mice measured by flow cytometry. (B) Representative flow cytometry plots of the IFN-γ versus IL-17 staining. Cells were gated on lymphocytes (based on forward and side scatter), CD45, and CD4. n=4 mice per group. Representative of two independent experiments. **p≤0.01. Abbreviations: MLN, mesenteric lymph node; PPs, Peyer’s patches; SILP, small intestinal lamina propria; CoLP, colonic lamina propria; IEL, intraepithelial lymphocyte; IFN-γ, interferon gamma; Apc, adenomatous polyposis coli; IL-17, interleukin 17.

Article Snippet: After 72 hr, supernatants were collected and cells were rested for 24 hr with IL-2 (10 ng/ml) before staining as described below. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antigen Clone Conjugate Raised In Isotype Supplier Catalog Number Dilution/Concentration Purpose CD3e 145-2C11 NA Armenian hamster IgG1, κ BD Biosciences 553058 1:500 Cell stimulation CD28 37.51 NA Syrian hamster IgG2, λ 1 BD Biosciences 553294 1:2000 Cell stimulation CD45 30-F11 PerCP-Cy5.5 Rat IgG2b, κ BD Biosciences 561869 1:400 Flow cytometry CD4 RM4-5 Allophycocyanin Rat IgG1a, κ BD Biosciences 553051 1:100 Flow cytometry CD8a 53-6.7 Allophycocyanin Rat IgG1a, κ BD Biosciences 553035 1:100 Flow cytometry IFN-γ XMG1.1 FITC Rat IgG1, κ BD Biosciences 554411 1:100 Flow cytometry IL-17 TC11-18H10 PE Rat IgG1, κ BD Biosciences 559502 1:200 Flow cytometry IL-4 11B11 NA Rat IgG1, κ BioLegend 504107 10 µg/ml Cell polarization IFN-γ XMG1.2 NA Rat IgG1, κ BioLegend 505811 10 µg/ml Cell polarization IFN-γ XMG1.2 Biotin Rat IgG1, κ BioLegend 505803 1 µg/ml ELISA, 1:100 IF IF and ELISA IFN-γ R4-6A2 NA Rat IgG1, κ BioLegend 505704 0.5 µg/ml ELISA IL-17A TC11-18H10.1 NA Rat IgG1, κ BioLegend 506901 1 µg/ml ELISA IL-17A TC11-8H4 Biotin Rat IgG1, κ BioLegend 507001 0.5 µg/ml ELISA Granzyme B NGZB PE Rat IgG1a, κ eBioscience 12-8898 1:100 Flow cytometry Rat Ig Polyclonal Biotin Donkey Polyclonal Jackson ImmunoResearch 112-065-003 1:400 IF Streptavidin NA Horseradish peroxidase NA NA Jackson ImmunoResearch 016-030-084 1:1000 ELISA Streptavidin NA Alexa Fluor 488 NA NA Thermo Fisher Scientific (Carlsbad, CA) S-32354 1:400 IF CD8a 53-6.7 NA Rat IgG1a, χ Santa Cruz sc-18913 1:500 IF Open in a separate window Abbreviations: IFN-γ, interferon gamma; IL, interleukin; PE, phycoerythrin; IF, immunofluorescence; NA, not applicable.

Techniques: Flow Cytometry, Staining