Fluorescent probes and kits for differential diagnosis of rhabdomyosarcoma and neuroblastoma bone marrow metastasis and leukemia by flow cytometry
A neuroblastoma and flow cytometry technology, which is applied in the field of medical detection reagents, can solve the problems of rare reports on the diagnosis and differential diagnosis of lamina sarcoma and neuroblastoma, and achieve good clinical application prospects and high specificity. Effect
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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, item 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 prote...
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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