Tumor cell lines, PBMCs, and CD3+ T cells were separately incubated with serially diluted indicated concentrations (0.0001–100 µg/mL) of anti-B7-H3 mAb (10-2#c) or αB7-H3/CD3 for 1 h on ice and stained with APC-anti-human IgG (409306, BioLegend, San Diego, CA, USA) in the dark for 30 min. Cell binding activity was analyzed using flow cytometry (CytoFLEX LX, Beckman Coulter, Brea, CA, USA).
Apc anti human igg
Manufactured by BioLegend
Sourced in United States
APC anti-human IgG is a labelled antibody used for flow cytometric analysis. It specifically binds to human immunoglobulin G (IgG) molecules. The APC (Allophycocyanin) fluorescent dye is conjugated to the antibody, allowing for the detection and quantification of IgG-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Cytoflex lx, by Beckman Coulter (1 mentions)
Ab171870, by Abcam (1 mentions)
Accuri c6 cytometer, by BD (1 mentions)
Apc anti human cd24 antibody, by BioLegend (1 mentions)
Apc anti human epcam, by BioLegend (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Mg132, by Merck Group (1 mentions)
Monoclonal antibody against β actin, by Cell Signaling Technology (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Protein a g plus agarose, by Merck Group (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Apc annexin 5 apoptosis detection kit, by BioLegend (1 mentions)
Ubiquitin, by Cell Signaling Technology (1 mentions)
Abt 263, by Santa Cruz Biotechnology (1 mentions)
Facsaria 3, by BD (1 mentions)
Anti human cd14 apc cy7, by BioLegend (1 mentions)
4 protocols using apc anti human igg
1
Cell Binding Assay for Anti-B7-H3 mAb
2
Isolation and Characterization of LGR5+ Glioma Cells
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Parent cells and enriched cells were incubated with LGR5 (#ab75735; Abcam) and an isotype control (#ab171870; Abcam) and subsequently with secondary antibodies (Alexa Fluor 488) for 30 min at room temperature and washed three times in phosphate buffered saline (PBS) after each reaction to detect the proportion of LGR5 expression. Using the method described above, LGR5+ and LGR5− glioma cells were sorted from parent cells by FACS.
To explore the proportion of reported stem cell markers in both LGR5+ and LGR5− glioma cells, the cells were incubated with APC anti-human CD133 antibody (#130–090-826; Miltenyi), APC anti-human CD44 antibody (#17–0441-82; eBioscience), APC anti-human CD24 antibody (#311118; BioLegend), APC anti-human CD90 antibody (#328114; BioLegend) or APC anti-human EpCAM (#328208; BioLegend) antibody for 30 min at room temperature. APC anti-human IgG (#409306; BioLegend) was used as an isotype control for these markers. All stained cell suspensions were detected or sorted using an AccuriC6 cytometer (BD Biosciences) and analyzed by FlowJo 7.6.1.
To explore the proportion of reported stem cell markers in both LGR5+ and LGR5− glioma cells, the cells were incubated with APC anti-human CD133 antibody (#130–090-826; Miltenyi), APC anti-human CD44 antibody (#17–0441-82; eBioscience), APC anti-human CD24 antibody (#311118; BioLegend), APC anti-human CD90 antibody (#328114; BioLegend) or APC anti-human EpCAM (#328208; BioLegend) antibody for 30 min at room temperature. APC anti-human IgG (#409306; BioLegend) was used as an isotype control for these markers. All stained cell suspensions were detected or sorted using an AccuriC6 cytometer (BD Biosciences) and analyzed by FlowJo 7.6.1.
3
Apoptosis Signaling Pathway Analysis
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ACF was purchased from Sigma-Aldrich; Merck KGaA. APC Annexin V Apoptosis Detection Kit was purchased from BioLegend, Inc (cat. no. 640930). APC anti-human IgG was purchased from BioLegend, Inc. (cat. no. 366906). ABT-263 was purchased from Santa Cruz Biotechnology, Inc. (cat. no. sc-207241). Monoclonal antibodies to detect MCL-1 (product no. 5453), GSK-3β (product no. 9315), phosphorylated (p)-GSK-3β (product no. 9323), GAPDH (product no. 2118) and polyclonal antibodies to detect p-MCL1 (product no. 4579), BCL-XL (product no. 2762), BCL-2 (product no. 2872), β-catenin (product no. 9562), cleaved PARP (product no. 9541), caspase-8 (product no. 9746), caspase-9 (product no. 9502), ubiquitin (product no. 3933), XAF1 (product no. 13805S) and XIAP (product no. 2042) were purchased from Cell Signaling Technology, Inc. Monoclonal antibody against β-actin (cat. no. A5316) was obtained from Sigma-Aldrich; Merck KGaA. Monoclonal antibodies to evaluate caspase-3 activation were purchased from Abcam (product code ab136812). MG-132 (product no. M7449), and cycloheximide (CHX; product no. C4859) were purchased from Sigma-Aldrich; Merck KGaA. Protein A/G PLUS-Agarose (cat. no. sc-2003) was purchased from Santa Cruz Biotechnology, Inc.
4
Identification of HIV Antibody-Producing Cells
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PBMC was purified by ficoll gradient from HIV infected patient, incubated with 1 μg/mlbiotinylated SF162gp140 trimer at 4°C for 30 min. The PBMC was then washed by PBS and stained with second antibodies:Pacific Blue™ anti-human CD19 (Biolegend, 302224), PE/Cy7 anti-human CD3(Biolegend, 300316), APC/Cy7 anti-human CD14 (Biolegend, 325620), APC anti-human IgG(Biolegend, 409306), 7-AAD(7-Aminoactinomycin D) (Invitrogen, 302232), FITC anti-human CD27(Biolegend, 302806), PerCP/Cy5.5 anti-human IgM (Biolegend, 314512), and R-Phycoerythrin Streptavidin (Jackson immunolab, 016-110-084). SF162gp140-trimerbinding memory B cells (CD19+, IgG+, SF162gp140+7AAD-IgM-CD14-CD3-) were sorted in 96 well PCR plate by FACS AriaIII(BD). The sorted cells were then lysedwith RNA directly reverse transcribed into cDNA according SuperScript™ III CellsDirectcDNA Synthesis System (Invitrogen, 18080-300) manual. Single cell derived cDNA was used for antibody variable gene amplification.
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