Combined immunoperoxidase analysis for visualization of cells of the blood-brain barrier

Bull Exp Biol Med. 2006 Oct;142(4):507-10. doi: 10.1007/s10517-006-0404-9.
[Article in English, Russian]

Abstract

A method for double immunoperoxidase staining of the blood-brain barrier cell elements was developed using 2mB6 monoclonal antibodies specifically visualizing brain capillary antigen and antibodies to glial fibrillary acidic protein (GFAP). The method is based on consecutive visualization of antigen structures in one section: first using a substrate mixture containing diaminobenzidine (brown coloring) and then a mixture with diaminobenzidine and CoCl(2) (blue coloring). This method visualizes cerebral capillary cells and fibrillar astrocytes interacting with them. The method can be used in all variants of double immunoperoxidase studies and for immunodetection of several antigens in immunoblotting analysis.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Astrocytes / cytology
  • Blood-Brain Barrier*
  • Capillaries / cytology*
  • Cerebrovascular Circulation / physiology*
  • Endothelium, Vascular / cytology
  • Glial Fibrillary Acidic Protein / analysis
  • Immunoenzyme Techniques*
  • Male
  • Rats
  • Rats, Wistar

Substances

  • Antibodies, Monoclonal
  • Glial Fibrillary Acidic Protein