Fluorescent probe and kit for carrying out differential diagnosis on leukemia and bone marrow metastasis of rhabdomyosarcoma and neuroblastoma by using flow cytometry
A technology for neuroblastoma and rhabdomyosarcoma, which is applied in measuring devices, instruments, and individual particle analysis, can solve the problems of rare reports on the diagnosis and differential diagnosis of stroma and neuroblastoma, and achieve good clinical application prospects , highly specific effects
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Embodiment 1
[0043] GD2-FITC (self-prepared, BALB / c mice were immunized with neuroblastoma cell line LAN-1, mouse B lymphocytes were fused with mouse myeloma cell line NS1 to form hybridoma cells to prepare monoclonal antibody GD2 and labeled For FITC fluorescein, refer to the magazine CANCER RESEARCH for specific technical steps, page 2988, Issue 46, 1986; Highly selective recognition of human neuroblastoma cells by mouse monoclonal antibody to a cytoplasmic antigen), CD90-PE (BD Company of the United States, article number: 555596), CD45 -PerCP (BD Company of the United States, article number: 347464), CD56-APC (BD Company of the United States, article number: 341025).
[0044] 1. Kit composition:
[0045] 1) Probe solution (2mL×1 bottle):
[0046] GD2-FITC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with fluorescein isothiocyanate (50μg / mL);
[0047] CD90-PE: 2mL PBS (containing 0.1% sodium azide) containing phycoerythrin-labeled 100μg antibody pr...
Embodiment 2
[0071] 1. Kit composition:
[0072] 1) Probe solution (2mL×1 bottle):
[0073] GD2-FITC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with fluorescein isothiocyanate (50μg / mL);
[0074] CD90-PE: 2mL PBS (containing 0.1% sodium azide) containing phycoerythrin-labeled 100μg antibody protein (50μg / mL);
[0075] CD45-PerCP: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with paramecium chlorophyll protein (50μg / mL);
[0076] CD56-APC: 2mL PBS (containing 0.1% sodium azide) containing 100μg antibody protein labeled with isophycocyanin (50μg / mL);
[0077] The antibody concentration ratio of the composition GD2-FITC: CD90-PE: CD45-PerCP: CD56-APC is 1:1:1:1.
[0078] 2) 10×hemolytic agent (100mL×1 bottle): 8.3% ammonium chloride, containing 15% paraformaldehyde;
[0079] 3) 10× phosphate buffer saline (PBS, pH=7.4, 1M, containing 1% sodium azide) (100mL×1 bottle);
[0080] 4) Positive standard (2mL×1 bottle): ...
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