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Immunofluorescent Staining of Murine Tissue

Zheng Liu Zheng Liu

Published: May 5, 2011 DOI: 10.21769/BioProtoc.65 Views: 17277

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Abstract

Renal immune complex deposition and leukocyte infiltration are characteristic of lupus nephritis in human patients and lupus-prone mice. This protocol describes how to stain frozen sections of murine kidney to study these features using fluorescent microscopy. This protocol was developed or modified in Dr. Anne Davidson’s lab at Feinstein Institute for Medical Research.

Keywords: Lupus search Nephrtitis search Mouse search Immunofluorescent search Frozen search

Materials and Reagents

  1. Antibodies
    1. Rat anti-mouse IgG2a FITC (Southern Biotech, catalog number: 1155-02 )
    2. Rat anti-mouse IgG3 FITC (Southern Biotech, catalog number: 1190-02 )
    3. Rat anti-mouse IgD PE (Southern Biotech, catalog number: 1120-09 )
    4. Rat anti-mouse F480 PE (Life Technologies, Invitrogen™, catalog number: MF48004 )
    5. Rat anti-mouse B220 PE (BD Biosciences, Pharmingen™, catalog number: BD553090 )
    6. Hamster anti-mouse CD11c PE (BD Biosciences, Pharmingen™, catalog number: BD553802 )
    7. Rat anti-mouse CD4 PE (BD Biosciences, Pharmingen™, catalog number: BD553049 )
      Note: The above antibodies have been tested by the author and may be substituted with the antibodies desired by users.

  2. Other materials
    1. Murine kidney tissues
    2. Acetone (Sigma- Aldrich, catalog number: 650501-4L )
    3. Tek O.C.T. Compound (Sakura Finetek, catalog number: 4587 )
    4. 3% Fetal bovine serum (FBS) (Sigma- Aldrich, catalog number: F2442-500ML ) in PBS.
    5. 0.5% Mouse BD Fc Block (BD Biosciences, Pharmingen™, catalog number: 553141 )
    6. DAPI nucleic acid stain (Life Technologies, Invitrogen™, catalog number: D1306 )
    7. Glycergel mounting medium (Dako, catalog number: C0563 )
    8. Block solution

Equipment

  1. Glass staining jars (Cole-Parmer, catalog number: EW-48585-00 )
  2. ImmEdge Pen (Vector Laboratories, Inc, catalog number: #H-4000 )

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Immune cell staining > Immunodetection

Immunology > Immune cell function > General

Cell Biology > Cell imaging > Fluorescence

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