实验方案(Protocols)

B Cell activation

Summary

B cells are activated and proliferate upon activation by specific antigens, anti-Ig antibodies, and/or mitogens.

Principle

B The basic principle of cell activation is that activation and proliferation can occur following activation by specific antigens, anti-Ig antibodies, and/or mitogens.

Operation method

B Cell activation

Materials and Instruments

Reagents:
(1) Purified B cells.
(2) Anti-IgM antibody, bacterial LPS (E. coli; Difco #011B4).
(3) Soluble CD40L, anti-mouse or human CD40 antibody.
(4) RPMI 1640 culture medium containing 10% FCS.
Instrumentation:
(1) 48-well flat-ground culture plate.
(2) Flow cytometer

Move

The basic process of B cell activation can be divided into the following steps:


(1) Purified B cells, adjust the B cell concentration to 10/ml with RPMI 1640 culture medium containing 10% FCS.


(2) Add 1 ml of B cell suspension to a 48-well flat-bottom plate, i.e. 1x10/well.


(3) Add one of the following stimuli to the B cell suspension and set up 3 replicate wells for each condition:


(1) Anti-IgM antibody, 2~200μg/ml;


2) LPS, 1~100 μg/ml.


3) sCD40L, 0.03~0.1μg/ml.


4) anti-CD40 antibody, 0.2~0.5μg/ml;


(5) negative control without any stimulant.


(4) Place the culture plate in a 37℃, 5% CO2 cell culture incubator for 24-72 hours to obtain activated B cells.


(5) Collect the B cells and surface label the activated B cells with FACS method for the expression of surface molecules, such as MHC class II molecules, CD80 and CD86.



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目录: 实验方案(Protocols)
探索主题: Immunological experiments

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阿拉丁科学.《B Cell activation》. 阿拉丁知识库,更新于 2026年7月24日。 https://www.aladdin-e.com/zh_cn/faqs/b-cell-activation-en.html
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