fluorochrome conjugated mouse anti-sheep cd4-fitc  (Bio-Rad)

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: CD4 + and CD8 + T cells in PBMCs. Peripheral blood samples were collected at different immunization times, and CD4 + and CD8 + T cells in PBMCs were analyzed using flow cytometry. (A) Proportion of CD4 + and CD8 + T cells in PBMCs of each group at week 8. (B, C) show the trend of the proportion of CD4 + and CD8 + T cells at different times, respectively. (D) The ratio of CD4 + to CD8 + T cells in each group of PBMCs at week 8. (E) The trend of the ratio of CD4 + to CD8 + T cells at different times. Data were obtained from 9 sheep, and results are presented as mean ± SD ( ns , P > 0.05).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in PBMCs by flow cytometry. PBMCs were stimulated with rEg.P29, and the cells were collected and labeled with antibodies to detect IFN-γ and IL-17A production by CD4 + and CD8 + T cells using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, D) show the IFN-γ production by CD4 + and CD8 + T cells in each group at week 8, respectively. (C, E) show the tendency of IFN-γ production by CD4 + and CD8 + T cells at different times, respectively. (F) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (G, I) show the IL-17A production by CD4 + and CD8 + T cells in each group of samples at week 8, respectively. (H, J) show the tendency of IL-17A production by CD4 + and CD8 + T cells at different times, respectively. Data were obtained from 9 sheep, and results are presented as mean ± SD (* P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in spleen lymphocytes by flow cytometry. Spleen lymphocytes were obtained after euthanasia of sheep, cells were labeled with antibodies after rEg.P29 stimulation, and IFN-γ and IL-17A production by CD4 + and CD8 + T cells was detected using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, C) show the IFN-γ production by CD4 + and CD8 + T cells in each group, respectively. (D) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (E, F) show the IL-17A production by CD4 + and CD8 + T cells in each group, respectively. Data were obtained from 7 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Cytokines produced in lymphocytes of mesenteric lymph nodes by flow cytometry. Lymphocytes of mesenteric lymph nodes were obtained after euthanasia of sheep, cells were labeled with antibodies after rEg.P29 stimulation, and IFN-γ, IL-4, and IL-17A production by CD4 + and CD8 + T cells was detected using flow cytometry. (A) Representative flow scatter plots for detecting IFN-γ production by CD4 + and CD8 + T cells. (B, C) show the IFN-γ production by CD4 + and CD8 + T cells in each group, respectively. (D) Representative flow scatter plots for detecting IL-17A production by CD4 + and CD8 + T cells. (E, F) show the IL-17A production by CD4 + and CD8 + T cells in each group, respectively. Data were obtained from 7 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Produced, Flow Cytometry, Labeling

Journal: Frontiers in Immunology

Article Title: Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep

doi: 10.3389/fimmu.2023.1243204

Figure Lengend Snippet: Proliferation of CD4 + and CD8 + T cells. PBMCs were labeled with CFSE, stimulated in vitro with rEg.P29, and the decrease in CFSE fluorescence intensity of labeled cells was detected using flow cytometry to assess the proliferation of CD4 + and CD8 + T cells. (A) Histogram plots of CFSE fluorescence of lymphocytes, CD4 + , and CD8 + T cells. (B–D) represent the proliferation frequencies of lymphocytes, CD4 + , and CD8 + T cells, respectively. Data were obtained from 5 sheep, and results are presented as mean ± SD (**** P < 0.0001).

Article Snippet: To determine the cellular phenotypes of PBMCs, cells were directly stained with mouse anti-sheep CD4 (Clone: 44.38) and mouse anti-sheep CD8 (Clone: 38.65) antibodies (Bio-Rad AbD Serotec, Munich, USA), and incubated at 4°C for 30 min in the dark, followed by two washes with washing buffer.

Techniques: Labeling, In Vitro, Fluorescence, Flow Cytometry

Journal: STAR Protocols

Article Title: Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

doi: 10.1016/j.xpro.2022.101688

Figure Lengend Snippet:

Article Snippet: Anti-CD4 (Clone 44.38; mouse IgG2a; 1:400 dilution) , Bio-Rad , Cat# MCA2213GA.

Techniques: Virus, Recombinant, Sterility, Staining, Cell Culture, Hood, Flow Cytometry, Software

Journal: STAR Protocols

Article Title: Multiparameter flow cytometry assay to analyze the pulmonary T cell profiles in the ovine model of respiratory syncytial virus infection

doi: 10.1016/j.xpro.2022.101688

Figure Lengend Snippet:

Article Snippet: Anti-CD4 (Clone 44.38; mouse IgG2a; 1:400 dilution) , Bio-Rad , Cat# MCA2213GA.

Techniques: Virus, Recombinant, Sterility, Staining, Cell Culture, Hood, Flow Cytometry, Software

Journal: Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

Article Title: Activated platelets and platelet-leukocyte aggregates in the equine systemic inflammatory response syndrome

doi: 10.1177/10406387221077969

Figure Lengend Snippet: Cytometric gating protocol. A. Platelet gate (P1) on the basis of the forward (FSC) and side scatter (SSC), threshold FSC 20,000. B. Single-parameter fluorescence histogram of the APC-conjugated polyclonal goat anti-mouse antibody as isotype-negative control for CD41/61 (FL-4: 640 nm, filter 675/25 nm). C. Single-parameter fluorescence histogram gated for APC-conjugated monoclonal mouse anti-sheep antibodies against CD41/61. D. Dotplot representing platelet expression of CD41/61 ( y -axis; FL-4; Q1-UL indicating positive events for CD41/61) and expression of CD11a/18 (leukocytes; x -axis; FL-1, 488 nm, filter 530/30 nm; Q1-LR indicating positive events for CD 11a/18); dual-labeled events positive for CD41/61 and CD11a/18 representing 3.5% platelet-leukocyte aggregates in Q1-UR.

Article Snippet: Activated platelets and PLAs were measured by dual-labeling with conjugated antibodies (all Bio-Rad, formerly AbD Serotec) as described., An allophycocyanin (APC)-conjugated (LYNX rapid antibody conjugation kit; Bio-Rad) monoclonal mouse anti-sheep antibody against CD41/61 with cross-reactivity against equine platelet CD41/61 , , was used as platelet marker (MCA1095GA, dilution 1:50 with modified HEPES/Tyrod buffer ).

Techniques: Fluorescence, Negative Control, Expressing, Labeling