The Experts below are selected from a list of 3 Experts worldwide ranked by ideXlab platform
Haruo Onda - One of the best experts on this subject based on the ideXlab platform.
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Production of recombinant human relaxin 3 in AtT20 Cells.
Regulatory peptides, 2003Co-Authors: Hideki Kizawa, Kazunori Nishi, Yoshihiro Ishibashi, Masataka Harada, Tsuneo Asano, Nobuhiro Suzuki, Shuji Hinuma, Yukio Fujisawa, Haruo OndaAbstract:Relaxin 3 has been reported recently as a member of the insulin/IGF/relaxin family. To clarify the function of relaxin 3, we prepared recombinant human relaxin 3 using a mouse adrenocorticotrophic hormone (ACTH)-Secreting Cell line, AtT20. To detect a mature form of recombinant human relaxin 3, a competitive enzyme immunoassay (EIA) was developed using a monoclonal antibody (mAb; HK4-144-10), which was raised for the N-terminal peptide of human relaxin 3 A-chain. We detected immunoreactive (ir-) relaxin 3 in the culture supernatant of AtT20 Cells stably transfected with human relaxin 3 cDNA. After treatment with 5 microM forskolin for 3 days, the concentration of the ir-relaxin 3 in the culture supernatant reached 12 nM. Ir-relaxin 3 was purified from the culture supernatant by a combination of various chromatographies. By analyses of N-terminal amino acid sequence and electrospray ionization mass spectrometry (ESI-MS), we confirmed that the purified material was a mature form of human relaxin 3. The recombinant human relaxin 3 thereby obtained increased intraCellular cAMP production in THP-1 Cells. Our results demonstrate that the expression of relaxin 3 cDNA in AtT20 Cells is a useful tool to produce a bioactive and mature form of relaxin 3.
Hideki Kizawa - One of the best experts on this subject based on the ideXlab platform.
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Production of recombinant human relaxin 3 in AtT20 Cells.
Regulatory peptides, 2003Co-Authors: Hideki Kizawa, Kazunori Nishi, Yoshihiro Ishibashi, Masataka Harada, Tsuneo Asano, Nobuhiro Suzuki, Shuji Hinuma, Yukio Fujisawa, Haruo OndaAbstract:Relaxin 3 has been reported recently as a member of the insulin/IGF/relaxin family. To clarify the function of relaxin 3, we prepared recombinant human relaxin 3 using a mouse adrenocorticotrophic hormone (ACTH)-Secreting Cell line, AtT20. To detect a mature form of recombinant human relaxin 3, a competitive enzyme immunoassay (EIA) was developed using a monoclonal antibody (mAb; HK4-144-10), which was raised for the N-terminal peptide of human relaxin 3 A-chain. We detected immunoreactive (ir-) relaxin 3 in the culture supernatant of AtT20 Cells stably transfected with human relaxin 3 cDNA. After treatment with 5 microM forskolin for 3 days, the concentration of the ir-relaxin 3 in the culture supernatant reached 12 nM. Ir-relaxin 3 was purified from the culture supernatant by a combination of various chromatographies. By analyses of N-terminal amino acid sequence and electrospray ionization mass spectrometry (ESI-MS), we confirmed that the purified material was a mature form of human relaxin 3. The recombinant human relaxin 3 thereby obtained increased intraCellular cAMP production in THP-1 Cells. Our results demonstrate that the expression of relaxin 3 cDNA in AtT20 Cells is a useful tool to produce a bioactive and mature form of relaxin 3.
Kazunori Nishi - One of the best experts on this subject based on the ideXlab platform.
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Production of recombinant human relaxin 3 in AtT20 Cells.
Regulatory peptides, 2003Co-Authors: Hideki Kizawa, Kazunori Nishi, Yoshihiro Ishibashi, Masataka Harada, Tsuneo Asano, Nobuhiro Suzuki, Shuji Hinuma, Yukio Fujisawa, Haruo OndaAbstract:Relaxin 3 has been reported recently as a member of the insulin/IGF/relaxin family. To clarify the function of relaxin 3, we prepared recombinant human relaxin 3 using a mouse adrenocorticotrophic hormone (ACTH)-Secreting Cell line, AtT20. To detect a mature form of recombinant human relaxin 3, a competitive enzyme immunoassay (EIA) was developed using a monoclonal antibody (mAb; HK4-144-10), which was raised for the N-terminal peptide of human relaxin 3 A-chain. We detected immunoreactive (ir-) relaxin 3 in the culture supernatant of AtT20 Cells stably transfected with human relaxin 3 cDNA. After treatment with 5 microM forskolin for 3 days, the concentration of the ir-relaxin 3 in the culture supernatant reached 12 nM. Ir-relaxin 3 was purified from the culture supernatant by a combination of various chromatographies. By analyses of N-terminal amino acid sequence and electrospray ionization mass spectrometry (ESI-MS), we confirmed that the purified material was a mature form of human relaxin 3. The recombinant human relaxin 3 thereby obtained increased intraCellular cAMP production in THP-1 Cells. Our results demonstrate that the expression of relaxin 3 cDNA in AtT20 Cells is a useful tool to produce a bioactive and mature form of relaxin 3.
Yoshihiro Ishibashi - One of the best experts on this subject based on the ideXlab platform.
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Production of recombinant human relaxin 3 in AtT20 Cells.
Regulatory peptides, 2003Co-Authors: Hideki Kizawa, Kazunori Nishi, Yoshihiro Ishibashi, Masataka Harada, Tsuneo Asano, Nobuhiro Suzuki, Shuji Hinuma, Yukio Fujisawa, Haruo OndaAbstract:Relaxin 3 has been reported recently as a member of the insulin/IGF/relaxin family. To clarify the function of relaxin 3, we prepared recombinant human relaxin 3 using a mouse adrenocorticotrophic hormone (ACTH)-Secreting Cell line, AtT20. To detect a mature form of recombinant human relaxin 3, a competitive enzyme immunoassay (EIA) was developed using a monoclonal antibody (mAb; HK4-144-10), which was raised for the N-terminal peptide of human relaxin 3 A-chain. We detected immunoreactive (ir-) relaxin 3 in the culture supernatant of AtT20 Cells stably transfected with human relaxin 3 cDNA. After treatment with 5 microM forskolin for 3 days, the concentration of the ir-relaxin 3 in the culture supernatant reached 12 nM. Ir-relaxin 3 was purified from the culture supernatant by a combination of various chromatographies. By analyses of N-terminal amino acid sequence and electrospray ionization mass spectrometry (ESI-MS), we confirmed that the purified material was a mature form of human relaxin 3. The recombinant human relaxin 3 thereby obtained increased intraCellular cAMP production in THP-1 Cells. Our results demonstrate that the expression of relaxin 3 cDNA in AtT20 Cells is a useful tool to produce a bioactive and mature form of relaxin 3.
Masataka Harada - One of the best experts on this subject based on the ideXlab platform.
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Production of recombinant human relaxin 3 in AtT20 Cells.
Regulatory peptides, 2003Co-Authors: Hideki Kizawa, Kazunori Nishi, Yoshihiro Ishibashi, Masataka Harada, Tsuneo Asano, Nobuhiro Suzuki, Shuji Hinuma, Yukio Fujisawa, Haruo OndaAbstract:Relaxin 3 has been reported recently as a member of the insulin/IGF/relaxin family. To clarify the function of relaxin 3, we prepared recombinant human relaxin 3 using a mouse adrenocorticotrophic hormone (ACTH)-Secreting Cell line, AtT20. To detect a mature form of recombinant human relaxin 3, a competitive enzyme immunoassay (EIA) was developed using a monoclonal antibody (mAb; HK4-144-10), which was raised for the N-terminal peptide of human relaxin 3 A-chain. We detected immunoreactive (ir-) relaxin 3 in the culture supernatant of AtT20 Cells stably transfected with human relaxin 3 cDNA. After treatment with 5 microM forskolin for 3 days, the concentration of the ir-relaxin 3 in the culture supernatant reached 12 nM. Ir-relaxin 3 was purified from the culture supernatant by a combination of various chromatographies. By analyses of N-terminal amino acid sequence and electrospray ionization mass spectrometry (ESI-MS), we confirmed that the purified material was a mature form of human relaxin 3. The recombinant human relaxin 3 thereby obtained increased intraCellular cAMP production in THP-1 Cells. Our results demonstrate that the expression of relaxin 3 cDNA in AtT20 Cells is a useful tool to produce a bioactive and mature form of relaxin 3.
