apc-conjugated anti-human cd47  (Thermo Fisher)

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Characteristics of the 24 patients treated at the National Cancer Center Hospital between 2003 and 2021.

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques:

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Immunohistochemical expression of CD47 in biopsy samples. The depicted regions were selected from incisional biopsy samples, and immunohistochemical staining was performed as described under Materials and Methods. The upper panel (A, from case 14) depicts CD47-negative expression, while the lower panel (B, from case 15) depicts CD47-positive expression, showing CD47-positive cells stained in light brown color.

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Comparison of clinicopathological characteristics based on CD47 positivity.

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: Comparison, Biomarker Discovery

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: In vitro efficacy of CD47 antibody in osteosarcoma cells. ( A ) The expression of CD47 is analyzed using FACS analysis in osteosarcoma cells. Representative images display the histogram of CD47 expression under basal conditions. ( B – D ) Osteosarcoma cells are treated with either isotype control or anti-CD47 (B6H12) antibody at the indicated concentrations for 24, 48, and 72 h. The effect of the anti-CD47 antibody on osteosarcoma cell viability is assessed using the EZ-CyTox cell viability assay kit. ( E ) KHOS/NP osteosarcoma cells are exposed to an isotype control or anti-CD47 (B6H12) antibody at the indicated concentrations for 24, 48, and 72 h. The effect of the anti-CD47 antibody on KHOS/NP cell apoptosis is evaluated using the Annexin V Detection kit.

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: In Vitro, Expressing, Control, Viability Assay

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Wound-healing assay to assess migration of osteosarcoma cells following CD47 (B6H12) antibody treatment. ( A ) MG-63 osteosarcoma cells are treated with a 10 μg/mL concentration of an isotype control antibody or the anti-CD47 antibody (B6H12). Representative images show the area the cells cover at the indicated time points after in vitro scratch wounding. ( B ) The summary graph illustrates the percentage of wound closure in MG-63 osteosarcoma cells over time compared with untreated controls. Statistical analysis of wound closure is conducted using ImageJ software with the wound healing size plug-in. The percentages are represented as the mean ± standard error of the mean, and statistical significance is determined by comparison with the control (* p < 0.05).

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: Wound Healing Assay, Migration, Concentration Assay, Control, In Vitro, Software, Comparison

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Wound-healing assay to assess migration of osteosarcoma cells following CD47 siRNA transfection. ( A ) MG-63 osteosarcoma cells are treated with either control siRNA or CD47 siRNA, and Western blot analysis is performed to evaluate CD47 expression. The summary graph illustrates the CD47 expression levels in the MG-63 osteosarcoma cells. ( B ) Representative images display the area covered by MG-63 osteosarcoma cells at the indicated time points following in vitro scratch wounding. The summary graph illustrates the percentage of wound closure in MG-63 cells over time compared to the control siRNA. Statistical analysis of wound closure is performed using ImageJ software with the wound healing size plug-in. The percentages are represented as the mean ± standard error of the mean, and statistical significance is determined by comparison with the control siRNA (** p < 0.01, n.s. : no significance).

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: Wound Healing Assay, Migration, Transfection, Control, Western Blot, Expressing, In Vitro, Software, Comparison

Journal: Cells

Article Title: CD47 in Osteosarcoma: Correlation with Metastasis and Macrophage-Mediated Phagocytosis

doi: 10.3390/cells13221862

Figure Lengend Snippet: Phenotypic characterization of monocyte-derived macrophages and comparative analysis of in vitro macrophage-mediated phagocytosis following CD47 (B6-H12) antibody treatment. ( A ) CD14-positive monocytes are isolated and cultured in a macrophage medium, followed by activation with recombinant human M-CSF for 6 d. Single cells are stained and analyzed using the BD FACSVerse System. ( B , C ) KHOS/NP cells are labeled with CellTrace™ CFSE dye (KHOS-CFSE) and incubated with 10 μg/mL anti-CD47 antibody or IgG ISO-type antibody for 30 min at 37 °C. KHOS/NP-CFSE cells are co-cultured with differentiated macrophages for 2 h. The percentage of macrophage-mediated phagocytosis (CD14-high, CellTrace-high, and CD11b-high) is measured by flow cytometry. Data are recorded using BD FACSVerse and analyzed using FlowJo software. The percentages are presented as the mean ± standard error of the mean, and statistical significance is determined by comparison with the IgG ISO-type antibody (** p < 0.01).

Article Snippet: Immunohistochemical staining was performed using allophycocyanin (APC)-conjugated anti-human CD47 (B6H12; Thermo Fisher Scientific, Waltham, MA, USA) and APC-conjugated mouse IgG1κ isotype control (MOPC-21; BioLegend, San Diego, CA, USA) antibodies.

Techniques: Derivative Assay, In Vitro, Isolation, Cell Culture, Activation Assay, Recombinant, Staining, Labeling, Incubation, Flow Cytometry, Software, Comparison