A sensitive and reliable quantification method for mouse interleukin-12 p70 based on fluorometric sandwich ELISA (FS-ELISA).
Interleukin-12 (IL-12) p70 is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation; it is a heterodimer of p35 and p40 subunits. Here, we report a new, highly sensitive, and reliable method that employs fluorometric sandwich ELISA for quantification of the mouse IL-12 (moIL-12) p70 protein. Our method could quantify moIL-12 p70 in the range of approximately 0.5 to 500 pg/ml. In the assay, no signals were produced by the moIL-12 p40 monomer, homodimer [(p40)2], or mouse IL-23 even up to a concentration of 500 pg/ml; this ensures that our assay has a high specificity for moIL-12 p70. To demonstrate that our method can determine natural moIL-12 in real physiological/pathological samples, we monitored the moIL-12 p70 secretion from peritoneal exudative cells after lipopolysaccharide (LPS) stimulation. IL-12 p70 secretion as early as 3h after LPS stimulation was reliably detected due to the high sensitivity of the method.
Nakamura
Tadashi
T
Department of Biological and Chemical Engineering, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.
Kimura
Hirokazu
H
Kato
Masahiko
M
Kurashige
Satonori
S
Wakamatsu
Kaori
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003470:Culture Media; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005470:Fluorometry; D018664:Interleukin-12 / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008863:Microspheres; D010537:Peritoneum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010844:Picibanil / Q000494:pharmacology; D011933:Reagent Kits, Diagnostic; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D003470:Culture Media; D008070:Lipopolysaccharides; D011933:Reagent Kits, Diagnostic; D018664:Interleukin-12; D010844:Picibanil