TY - JOUR
T1 - Establishment of the anti-klotho monoclonal antibodies and detection of klotho protein in kidneys
AU - Kato, Yukinari
AU - Arakawa, Emi
AU - Kinoshita, Satoko
AU - Shirai, Akio
AU - Furuya, Akiko
AU - Yamano, Kazuya
AU - Nakamura, Kazuyasu
AU - Iida, Akihiro
AU - Anazawa, Hideharu
AU - Koh, Noritoshi
AU - Iwano, Akiko
AU - Imura, Akihiro
AU - Fujimori, Toshihiko
AU - Kuro-O, Makoto
AU - Hanai, Nobuo
AU - Takeshige, Kazuhiko
AU - Nabeshima, Yo Ichi
PY - 2000/1/19
Y1 - 2000/1/19
N2 - A novel gene, klotho (kl), which is involved in the development of a syndrome resembling human aging in mice, was recently identified. The kl gene encodes a single-pass membrane protein whose extracellular domain carries homology to b-glucosidases. There also exists a splice variant of kl mRNA which encodes a putative secreted protein in both human and mouse. In this study, to characterize the physiological roles of Klotho protein, we established three monoclonal antibodies (mAbs) against the recombinant human Klotho protein. The mAbs are named KM2076 (rat IgG2a), KM2119 (rat IgG2b), and KM2365 (mouse IgG1). In Western blots, KM2076 and KM2119 specifically recognized a 130 kDa Klotho protein in the mouse and human kidney membrane fractions. To detect the human Klotho protein, the sandwich-type ELISA system with KM2076 and KM2365 was established. Using the ELISA system, we detected the human Klotho protein as low as 20 ng/ml in the supernatant of Chinese hamster ovary cells (CHO cells), introduced the human klotho gene. KM2076 and KM2119 specifically gave a positive staining by immunohistochemical staining in paraffin or frozen sections of the kidneys from wild-type mice but not in those from kl mice. Strong staining was observed especially in cortical renal tubules of the mouse kidney, where expression of klotho transcripts overlaps. KM2076 also showed a similar reaction pattern in the paraffin sections of rat and human kidneys. The mAbs established in this paper will serve as useful analytical, pathological, and diagnostic tools to disclose the role of Klotho protein in the suppression of a syndrome resembling human aging. (C) 2000 Academic Press.
AB - A novel gene, klotho (kl), which is involved in the development of a syndrome resembling human aging in mice, was recently identified. The kl gene encodes a single-pass membrane protein whose extracellular domain carries homology to b-glucosidases. There also exists a splice variant of kl mRNA which encodes a putative secreted protein in both human and mouse. In this study, to characterize the physiological roles of Klotho protein, we established three monoclonal antibodies (mAbs) against the recombinant human Klotho protein. The mAbs are named KM2076 (rat IgG2a), KM2119 (rat IgG2b), and KM2365 (mouse IgG1). In Western blots, KM2076 and KM2119 specifically recognized a 130 kDa Klotho protein in the mouse and human kidney membrane fractions. To detect the human Klotho protein, the sandwich-type ELISA system with KM2076 and KM2365 was established. Using the ELISA system, we detected the human Klotho protein as low as 20 ng/ml in the supernatant of Chinese hamster ovary cells (CHO cells), introduced the human klotho gene. KM2076 and KM2119 specifically gave a positive staining by immunohistochemical staining in paraffin or frozen sections of the kidneys from wild-type mice but not in those from kl mice. Strong staining was observed especially in cortical renal tubules of the mouse kidney, where expression of klotho transcripts overlaps. KM2076 also showed a similar reaction pattern in the paraffin sections of rat and human kidneys. The mAbs established in this paper will serve as useful analytical, pathological, and diagnostic tools to disclose the role of Klotho protein in the suppression of a syndrome resembling human aging. (C) 2000 Academic Press.
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U2 - 10.1006/bbrc.1999.2009
DO - 10.1006/bbrc.1999.2009
M3 - Article
C2 - 10631108
AN - SCOPUS:0034685014
SN - 0006-291X
VL - 267
SP - 597
EP - 602
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -