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Cell Isolation of Spleen Mononuclear Cells

脾脏单核细胞的分离

BW Benno Weigmann

发布: 2013年05月05日第3卷第9期 DOI: 10.21769/BioProtoc.689 浏览次数: 33187

评审: Lin Fang

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Abstract

This method allows you to isolate different subclass mononuclear cells, like B-cells, T cells, CD4+ and CD8+ T, from mouse spleen. By conjugating cells with specific antibodies and subsequently magnetic beads isolation, using the technique from Miltenyi, this allows a high purity.

Keywords: T cell (T细胞) search Purification (净化) search Lymphocyte (淋巴细胞) search Isolation (隔离) search

Materials and Reagents

  1. Antibody
    1. FITC-conjugated anti-CD3 antibody (BD Biosciences, catalog number: 553058 , clone 145-2C11)
    2. PE-conjugated anti-CD19 antibody (BD Biosciences, catalog number: 557399 , clone 1D3)
    3. FITC-conjugated anti-CD4 antibody (BD Biosciences, catalog number: 557667 , clone RM4-5)
    4. PE-conjugated anti-CD8 antibody (BD Biosciences, catalog number: 561095 , clone 53-6.7)

  2. Microbeads
    1. Anti-CD43 microbeads (Miltenyi Biotec, catalog number: 130-049-801 )
    2. Anti-CD90 microbeads (Miltenyi Biotec, catalog number: 130-091-376 )
    3. Anti-CD4 microbeads (Miltenyi Biotec, catalog number: 130-049-201 )
    4. Anti-CD8 microbeads (Miltenyi Biotec, catalog number: 130-091-112 )

  3. Others
    1. 1x phosphate-buffered saline (PBS) (pH 7.2) (Life Technologies, Gibco®, catalog number: 10010-031 )
    2. Bovine serum albumin (BSA) (Life Technologies, InvitrogenTM, catalog number: 15561-020 )
    3. EDTA (Sigma-Aldrich, catalog number: EDS-100G )
    4. Ice-cold separation buffer (see Recipes)

Equipment

  1. Scissors and forceps
  2. MiniMACS separation unit (Miltenyi Biotec, MiniMACS Separator, catalog number: 130-090-312 )
  3. Separation column (Miltenyi Biotec, separation column, Type MS, catalog number: 130-042-201 )
  4. Cell strainer (BD Biosciences, catalog number: 352360 )
  5. Flow cytometer (BD Biosciences, Coulter)
  6. Shaker

Procedure

English
中文翻译

文章信息

版权信息

© 2013 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Weigmann, B. (2013). Cell Isolation of Spleen Mononuclear Cells. Bio-protocol 3(9): e689. DOI: 10.21769/BioProtoc.689.

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分类

免疫学 > 免疫细胞分离 > 淋巴细胞

细胞生物学 > 细胞分离和培养 > 细胞分离

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