Sequencing:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Activity Assay:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Cell Culture:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Cloning:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Ligand Binding Assay:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Amplification:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Reverse Transcription:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Polymerase Chain Reaction:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
Synthesized:Article Title: Insulin resistance curative and method of screening the same
Article Snippet: .. The base sequence human PPARγ gene is described in the gene database GenBank as Accession No. X90563, and the amino acid sequence is described as SEQ; NO. 6. (1-1) Obtainment of Total RNA From Cell Line HepG2 In the same manner as in the method described in the aforementioned determination of PTP-1B inhibitory activity, and (1-1) Obtainment of total RNA from cell line HepG, human liver tumor-derived cell line HepG2 (American Type Culture Collection HB-8065) purchased from Dainippon Pharmaceutical Co., Ltd. was cultured, and the total RNA was extracted using a trizol reagent (Gibco Trizol Reagent, manufactured by Invitrogen) following the instructions attached thereto. (1-2) Cloning of cDNA to PPARγ Ligand Binding Site As a primer for amplification by reverse transcription polymerase chain reaction (hereinafter RT-PCR) method of cDNA on the PPARγ ligand binding site, the following two deoxyoligo nucleotides (primers No. 3 and No. 4) designed based on the human PPARγ gene sequence were chemically synthesized using Beckman Oligo 1000 (manufactured by Beckman). .. 5′-ggatccataatgccatcaggtttgggcgg-3′ (primer No. 3, SEQ; NO. 7) 5′-aagcttctagtacaagtccttgtagatctc-3′ (primer No. 4, SEQ; NO. 8) Using HepG2 total RNA obtained earlier as a template and the above-mentioned primers No. 3 and No. 4 as primers, PPARγ cDNA was amplified by RT-PCR method using Ready-To-Go RT-PCR Beads (manufactured by Amersham Pharmacia Biotech).
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