A precise detection kit for tumor immune cell subgroup typing
A detection kit and immune cell technology, applied in the field of cell biology detection, can solve the problems of complex experimental operation, high cost, and few types of metal pre-labeled antibodies, and achieve the effects of improving detection efficiency, saving samples, and accurate typing
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Embodiment 1
[0062] Example 1 Immune cell typing detection antibody panel
[0063] This example provides a set of antibody panels for the detection of immune cell typing. The antibody panel contains a total of 42 metal pre-labeled antibodies. The antibody panel can identify all T cell subsets (including T, Treg, central memoryT, effector T, effector memory CD4 + T, CD8 + T) as well as B cells, NK cells, granulocytes, monocyte-macrophage subsets, etc., and also covers immune checkpoints and cell proliferation indicators.
[0064] The antibody panel combined with mass spectrometry and flow cytometry technology can be used for the typing and detection of immune cells, and 42 antibodies use 42 channels.
[0065] Table 1 shows the antibody clone numbers, metal element labels and antibody-bound markers of the 42 metal-prelabeled antibodies. Among them, markers numbered 11, 15, 24, and 38 are intracellular proteins, and others are extracellular (cell surface) proteins.
[0066] Table 1 Antibo...
Embodiment 2
[0069] Example 2 Application of Antibody Panel in Immune Cell Typing of Tumor Patients
[0070] In this example, the antibody panel of Example 1 is used to detect the typing of immune cells in tumor patients, and the specific method is as follows:
[0071] 1. Prepare fresh peripheral blood from normal people and tumor patients, and extract PBMC from peripheral blood mononuclear cells.
[0072] 2. Divide the peripheral blood mononuclear cells PBMC extracted in step 1 into two groups, the control group and the experimental group, each group is about 3 × 10 6 Cells were resuspended in PBS to adjust the volume to 1 mL.
[0073] 3. Add cisplatin (Cell-ID Cisplatin, 201094, Fluidigm) at a final concentration of 5 μM, and stain at room temperature for 2 minutes.
[0074] 4. Add 2ml of Cell Staining Buffer (201068, Fluidigm) to terminate the reaction, and centrifuge at 500×g for 5 minutes at room temperature.
[0075] 5. Aspirate the supernatant, add 50 μL blocking solution (5 μl Fc ...
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