TY - JOUR
T1 - Generation and characterization of an IGFBP-7 antibody
T2 - Identification of 31kD IGFBP-7 in human biological fluids and Hs578T human breast cancer conditioned media
AU - Wilson, Elizabeth M.
AU - Oh, Youngman
AU - Rosenfeld, Ron G.
PY - 1997
Y1 - 1997
N2 - The cDNA encoding mac25 (IGFBP-7) was first derived from mRNA isolated from leptomeningial and senescent human mammary epithelial cells (1,2). The open reading frame was shown to predict a protein with homology to the amino terminus of the IGF binding proteins, (IGFBP)1-6. Studies in our laboratory have shown that baculovirus generated mac25 binds IGF-I and-II in a specific manner, leading to the renaming of mac25 as IGFBP-7 (3). Further studies at the cellular level, to identify the involvement of IGFBP-7 in IGF regulation and cell growth, require a specific antibody against the protein, which has yet to be identified in either cultured cells or in vivo. We have now generated three polyclonal antibodies against the purified baculovirus peptide and, by western immunoblots and immunoprecipitation, demonstrated the existence of a specific 31,000 dalton protein. It is a secreted protein, and can be identified in the conditioned media of Hs578T breast cancer cells, as well as in normal human urine, cerebrospinal fluid and amniotic fluid. Subsequent studies with these antibodies should help elucidate the physiological role(s) of this protein.
AB - The cDNA encoding mac25 (IGFBP-7) was first derived from mRNA isolated from leptomeningial and senescent human mammary epithelial cells (1,2). The open reading frame was shown to predict a protein with homology to the amino terminus of the IGF binding proteins, (IGFBP)1-6. Studies in our laboratory have shown that baculovirus generated mac25 binds IGF-I and-II in a specific manner, leading to the renaming of mac25 as IGFBP-7 (3). Further studies at the cellular level, to identify the involvement of IGFBP-7 in IGF regulation and cell growth, require a specific antibody against the protein, which has yet to be identified in either cultured cells or in vivo. We have now generated three polyclonal antibodies against the purified baculovirus peptide and, by western immunoblots and immunoprecipitation, demonstrated the existence of a specific 31,000 dalton protein. It is a secreted protein, and can be identified in the conditioned media of Hs578T breast cancer cells, as well as in normal human urine, cerebrospinal fluid and amniotic fluid. Subsequent studies with these antibodies should help elucidate the physiological role(s) of this protein.
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U2 - 10.1210/jcem.82.4.3983
DO - 10.1210/jcem.82.4.3983
M3 - Article
C2 - 9100611
AN - SCOPUS:0030904370
SN - 0021-972X
VL - 82
SP - 1301
EP - 1303
JO - Journal of Clinical Endocrinology and Metabolism
JF - Journal of Clinical Endocrinology and Metabolism
IS - 4
ER -