mouse igg2b pe cy7 southern biotech  (SouthernBiotech)

Journal: Molecular Cancer

Article Title: The Wnt/β-catenin pathway regulates growth and maintenance of colonospheres

doi: 10.1186/1476-4598-9-212

Figure Lengend Snippet: Spheroids/colonospheres display increased levels of CSC markers . A . Representative scattered plots of flow cytometry analyses of CD44 positive cells in different colon cancer cells: Figures in the left panel depict the parental colon cancer cells stained with PerCP-Cy5 mouse IgG2b (Isotype), whereas those in the right panel, show the cells treated with CD44 conjugated PerCP-Cy5 mouse anti-human antibodies. Values represent percentage of cells that are positive for CD44-PerCP-Cy5. B . Photomicrographs showing the integrity and viability of colonospheres (HCT-116) using acridine orange-ethidium bromide (AO-EtBr) staining and immunofluorescence staining depicting the membrane bound expression of putative CSC marker EpCAM (Upper Panel); and flow cytometric analysis showing the proportion of CD44 positive cells in HCT-116 colonospheres-derived cells. Corresponding isotype for CD44 antibodies is shown (Lower Panel).

Article Snippet: Mouse anti-human PerCP-Cy5 IgG2b (isotype) or CD44 conjugated antibodies (Santa Cruz Biotechnology Inc., Santa Cruz, CA) and mouse anti-human PE-Cy7 IgG2b (isotype) or CD44 conjugated antibodies (BD Pharmingen) were used for flow cytometry.

Techniques: Flow Cytometry, Staining, Immunofluorescence, Expressing, Marker, Derivative Assay

Journal: Molecular Cancer

Article Title: The Wnt/β-catenin pathway regulates growth and maintenance of colonospheres

doi: 10.1186/1476-4598-9-212

Figure Lengend Snippet: Downregulation of β-catenin results in reduction of CD44 positive cells in spheroids/colonospheres and inhibits regeneration of colonospheres . A . Flow cytometric analysis shows CD44 positive cells in HCT-116 colonospheres following 72 hrs of transfection with β-catenin siRNA or the vector; corresponding isotypes are also shown, values represent percentage of cells that are positive for CD44-PE-Cy7. B . Western blot showing downregulation of β-catenin and CD44 following 72 hrs of transfection of β-catenin-siRNA, compared to the vector-control in HCT-116 colonospheres-derived cells. C . Colonosphere formation assay shows decreased regeneration of colonospheres by β-catenin-knockdown HCT-116 colonospheres cells. The numbers represent percent of corresponding control normalized to β-actin. * p < 0.001, compared to the corresponding vector-treated controls

Article Snippet: Mouse anti-human PerCP-Cy5 IgG2b (isotype) or CD44 conjugated antibodies (Santa Cruz Biotechnology Inc., Santa Cruz, CA) and mouse anti-human PE-Cy7 IgG2b (isotype) or CD44 conjugated antibodies (BD Pharmingen) were used for flow cytometry.

Techniques: Transfection, Plasmid Preparation, Western Blot, Derivative Assay, Tube Formation 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: 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 IgG2b, PE-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1090-17.

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 IgG2b, PE-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1090-17.

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 IgG2b, PE-Cy7 (Goat IgG; 1:250 dilution) , SouthernBiotech , Cat# 1090-17.

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