Rat LH (rLH) and FSH (rFSH) were measured by sensitive and specific competi
tion ELISAs. The rat LH ELISA used rLH-I-9 coated plates, an antiserum agai
nst rLH and an antibody against rabbit IgG labeled with peroxidase. Using r
LH-RP-3 as a standard, rat LH was determined by binding of the anti-LH anti
body to rLH-I-9 coated plates. The sensitivity of the assay was 0.8 ng/mL.
Similarly, the rat FSH-ELISA used rFSH-I-8 coated plates, an antiserum agai
nst rFSH and an antibody against rabbit IgG labeled with peroxidase. Using
rFSH-RP-3 as a standard, the FSH-ELISA was also determined by binding of th
e anti-FSH antibody to rFSH-I-8 coated plates. The sensitivity of this assa
y was 1.25 ng/mL. Both rat LH and FSH ELISA assays are highly specific and
provide accurate determination of gonadotrophins in buffers, sera, cell cul
ture media, and anterior pituitary extracts. These assays were used for mon
itoring the gonadotrophin surge-attenuating factor (GnSAF) and inhibin acti
vities present in human follicular fluid (hFF). The 2 new ELISA procedures
have practical advantages (safety, convenience, economy) over the RIA metho
ds, and they perform as well as the RIA techniques at the same range of con
centrations. (C) 1999 by Elsevier Science Inc.