Immunoenzymatic assay that measures the expression of murine histocompatibility antigens in macrophages and lymphocytes

✍ Scribed by G. Fragoso; F. Valdez; Y. Rosenstein; T. Govezensky; C. Larralde; Dra. E. Sciutto

• Publisher: John Wiley and Sons
• Year: 1993
• Tongue: English
• Weight: 420 KB
• Volume: 7
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860070609

✦ Synopsis

Abstract

A convenient—simple, sensitive, rapid and reproducible—enzyme immunoassay to measure H‐2 particulated and solubilized cellular antigens is described. Cellular antigens were measured by ELISA through the binding of specific biotinylated antibodies and streptoavidin‐peroxidase conjugate to cells in suspension. Endogenous peroxidase activity of activated cells was inhibited by addition of sodium azide and H~2~O~2~ in acid conditions. The assay proved capable of distinguishing between two cell lines (EL–4/H–2^b^ and P815/H–2^d^) and even between the cells of three congenic mouse strains (BALB/B, H–2^b^, BALB/c, H–2^d^, and BALB/K, H‐2^k^) and was sensitive to as few as 2.5 × 10^4^ cells/well. Results were comparable to those obtained with FACS. An inhibition version of this assay was found to be very useful for the detection of H–2 antigens present in whole antigen cells extracts. © 1993 Wiley‐Liss, Inc.