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Kit for simultaneously detecting various cell subpopulations and functional changes and application of kit

A technology of cell subgroups and kits, applied in the field of kits for simultaneous detection of multiple cell subgroups and functional changes, to achieve broad application prospects, convenient use, and high application value

Pending Publication Date: 2021-12-31
BLUE ELEGANT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the existing flow detection technology, it is difficult to detect multiple cell subpopulations simultaneously in one detection

Method used

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  • Kit for simultaneously detecting various cell subpopulations and functional changes and application of kit
  • Kit for simultaneously detecting various cell subpopulations and functional changes and application of kit
  • Kit for simultaneously detecting various cell subpopulations and functional changes and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0155] This embodiment provides an antibody composition, which includes a surface antibody composition and an intracellular antibody composition.

[0156] The surface antibody composition includes a combination of at least two of Zombie NIR, CD45, Ly6G, CD11b, F4 / 80, Ly6c, NK1.1, CD3, CD4, CD25, CD8a, CD19, CD11c, CD80 or CD206;

[0157] The intrabody composition includes a combination of at least two of FoxP3, IL-10, Granzyme B, Perforin, or IFN-γ.

[0158] In parts by weight, the antibody composition includes 0.1 parts of Zombie NIR, 0.3 parts of CD45, 0.3 parts of Ly6G, 0.3 parts of CD11b, 0.6 parts of F4 / 80, 0.08 parts of Ly6c, 0.5 parts of NK1.1, 0.2 parts of CD3, 0.2 parts of CD4 0.15 copies, 0.1 copies of CD25, 0.3 copies of CD8a, 0.2 copies of CD19, 1 copy of CD11c, 2 copies of CD80, 1 copy of CD206, 0.5 copies of FoxP3, 2.5 copies of IL-10, 0.3 copies of Granzyme B, 5 copies of Perforin and 5 copies of IFN-γ .

[0159] The antibody composition of the present inventi...

Embodiment 2

[0161] This embodiment provides a kit for simultaneously detecting multiple cell subpopulations, the kit for simultaneously detecting multiple cell subpopulations includes the antibody composition in Example 1;

[0162] The kit for simultaneously detecting multiple cell subpopulations also includes a blocking agent, membrane permeation fluid and flow cytometry fluid.

Embodiment 3

[0164] In this embodiment, the kit for simultaneously detecting multiple cell subpopulations in Example 2 is used to detect changes in subpopulations of immune cells in peripheral blood of mice, including the following steps:

[0165] (1) Preparation of single cell suspension

[0166] 1. Immediately take 200 μL of fresh blood into an EP tube containing 200 μL of 3.8% sodium citrate anticoagulant, mix well, and centrifuge at 1500g for 15 minutes at 4°C;

[0167] 2. Add 1mL PBS to resuspend and wash the blood cells, centrifuge at 350g for 5min at 4°C, and discard the supernatant;

[0168] 3. Add 3mL ACK red blood cell lysate to each tube of 15mL centrifuge tube, gently pipette and shake, and lyse on ice for 2min;

[0169] 4. Add 2 times the volume of HBSS solution, mix gently, centrifuge at 350g at 4°C for 5min, and discard the supernatant;

[0170] 5. Resuspend the cell pellet.

[0171] (2) After blocking with a blocking agent, stain with a surface antibody composition, wash...

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Abstract

The invention provides a kit for simultaneously detecting various cell subpopulations and functional changes and application of the kit. The kit comprises an antibody composition, and the antibody composition comprises a surface antibody composition and an intracellular antibody composition. The surface antibody composition comprises a combination of at least two of Zombie NIR, CD45, Ly6G, CD11b, F4 / 80, Ly6c, NK1.1, CD3, CD4, CD25, CD8a, CD19, CD11c, CD80 and CD206. The intracellular antibody composition comprises a combination of at least two of FoxP3, IL-10, Granzyme B, Perforin and IFNgamma. The invention also provides a method for detecting the change of the immune cell subpopulations. The kit can be used for simultaneously detecting changes of a plurality of immune cell subgroups and functions, and the result is comprehensive and specific.

Description

technical field [0001] The invention belongs to the technical field of immune level detection, and in particular relates to a kit for simultaneously detecting various cell subgroups and functional changes and an application thereof. Background technique [0002] Flow cytometry is a technique for continuous detection of single cells or particles of similar cell size in a cell population, which can perform rapid, high-throughput, and accurate simultaneous detection of multiple parameters of cells or particles. At present, flow cytometry technology has been widely used in hematology, immunology, oncology and other scientific research or clinical related testing fields, and is one of the important detection methods. [0003] The occurrence of diseases is often accompanied by changes in the immune level of the body. For example, during the occurrence of tumors, the main immune cell subsets in the blood, spleen, and tumor tissue will change significantly, including CD4 T cells, CD...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N15/14
CPCG01N33/56966G01N15/1434G01N2469/10G01N2015/1006G01N15/01
Inventor 戚何妹陶铸容志恩亢中奎丁文婷秦莉陈小平
Owner BLUE ELEGANT BIOTECH CO LTD
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