apc cy7 anti mouse igg2a  (SouthernBiotech)

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: Antibody titration and validation All antibodies were titrated separately; here are a few examples to illustrate. We started with the manufacturer recommended dilution and made at least 6-point 1:2 dilution series plus unstained control. (A) WC1-IgG2a-FITC: recommended dilution 1:10 corresponds to 1 μg/mL. (B) CD45RO-IgG3-PE, the manufacturer does not provide the concentration corresponding to the recommended dilution 1:10. (C) CD25-IgG3: recommended dilution 1:100 corresponds to 10 μg/mL. (D) CD8-IgG1: recommended dilution 1:40 corresponds to 25 μg/mL. (E) IgG2a-APC-Cy7: recommended dilution 1:250 corresponds to 1 μg/mL. (F) IgG1-PE-Cy7: recommended dilution 1:40 corresponds to 5 μg/mL. Highlighted are the working dilutions that were defined for further use in the study.

Article Snippet: Anti-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

Techniques: Titration, Biomarker Discovery, Control, Concentration Assay

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: Staining procedure using an eight-step, seven-color assay

Article Snippet: Anti-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

Techniques: Staining, Blocking Assay

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-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

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: Antibody titration and validation All antibodies were titrated separately; here are a few examples to illustrate. We started with the manufacturer recommended dilution and made at least 6-point 1:2 dilution series plus unstained control. (A) WC1-IgG2a-FITC: recommended dilution 1:10 corresponds to 1 μg/mL. (B) CD45RO-IgG3-PE, the manufacturer does not provide the concentration corresponding to the recommended dilution 1:10. (C) CD25-IgG3: recommended dilution 1:100 corresponds to 10 μg/mL. (D) CD8-IgG1: recommended dilution 1:40 corresponds to 25 μg/mL. (E) IgG2a-APC-Cy7: recommended dilution 1:250 corresponds to 1 μg/mL. (F) IgG1-PE-Cy7: recommended dilution 1:40 corresponds to 5 μg/mL. Highlighted are the working dilutions that were defined for further use in the study.

Article Snippet: Anti-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

Techniques: Titration, Biomarker Discovery, Control, Concentration Assay

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: Staining procedure using an eight-step, seven-color assay

Article Snippet: Anti-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

Techniques: Staining, Blocking Assay

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-mouse IgG2a, APC-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1080-19.

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

Journal: Molecules

Article Title: Blockade of NF-κB Translocation and of RANKL/RANK Interaction Decreases the Frequency of Th2 and Th17 Cells Capable of IL-4 and IL-17 Production, Respectively, in a Mouse Model of Allergic Asthma

doi: 10.3390/molecules26113117

Figure Lengend Snippet: Effect of inhibitors of IKK (BMS-345541 (BMS)), NF-κB translocation (dehydroxymethylepoxyquinomicin (DHMEQ)) and RANK/RANK-L interaction (osteoprotegerin (OPG)), and glucocorticosteroid (methylprednisolone (MP)) administration on the number of IL-4- and IL-17-producing CD8 + T cells. The relative count ( A , D ) is expressed as a percentage of IL-4- or IL-17-producing cells within CD8 + T cells. The absolute count ( B , E ) represents the number of IL-4- or IL-17-producing CD8 + T cells per lung sample collected from individual mice. * p < 0.05, ** p < 0.01, *** p < 0.001. Examples of dot plot cytograms showing the distribution of IL-4- or IL-17-producing and non-producing cells within CD8 + T cells ( C and F , respectively).

Article Snippet: Subsequently, the cells were stained for surface antigens with fluorochrome-conjugated monoclonal antibodies (mAbs): PerCP-Cy 5.5 rat anti-mouse CD4 (clone RM4-5, IgG2a, κ), APC-Cy7 rat anti-mouse CD8a (clone 53-6.7, IgG2a, κ) and PE-Cy7 rat anti-mouse CD25 (clone PC61, IgG1, λ; all from BD Biosciences).

Techniques: Translocation Assay

Journal: Molecules

Article Title: Blockade of NF-κB Translocation and of RANKL/RANK Interaction Decreases the Frequency of Th2 and Th17 Cells Capable of IL-4 and IL-17 Production, Respectively, in a Mouse Model of Allergic Asthma

doi: 10.3390/molecules26113117

Figure Lengend Snippet: Gating strategy for flow cytometric data analysis and calculation of the absolute cell counts of lymphocyte subsets. Lymphocytes were identified based on forward and side scatter (FSC/SSC) properties, and then gated for expression of CD4 or CD8 surface receptors. CD4 + T cells were analyzed for expression/co-expression of CD25 and Foxp3. On this basis, Treg (Foxp3 + CD25 + CD4 + ) and non-Treg (the remaining CD4 + T cells, i.e., non-Foxp3 + CD25 + CD4 + T cells) cells were distinguished. Subsequently, IL-4-, IL-10-, IL-17- and TGF-β-producing cells were identified within particular cell subsets. Absolute cell counts of lymphocyte subsets (i.e., number of cells from particular subpopulations per mediastinal lymph node or lung sample) were calculated using the dual platform method, as shown above.

Article Snippet: Subsequently, the cells were stained for surface antigens with fluorochrome-conjugated monoclonal antibodies (mAbs): PerCP-Cy 5.5 rat anti-mouse CD4 (clone RM4-5, IgG2a, κ), APC-Cy7 rat anti-mouse CD8a (clone 53-6.7, IgG2a, κ) and PE-Cy7 rat anti-mouse CD25 (clone PC61, IgG1, λ; all from BD Biosciences).

Techniques: Expressing