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All Journal ACTA VETERINARIA INDONESIANA Jurnal Veteriner Indonesia Medicus Veterinus Farmaka Majalah Kedokteran Bandung Jurnal Kesehatan Masyarakat Althea Medical Journal Journal of Biomedicine and Translational Research Dharmakarya : Jurnal Aplikasi Ipteks Untuk Masyarakat Jurnal Kesehatan Andalas Folia Medica Indonesiana Indonesian Journal of Pharmaceutical Science and Technology Jurnal Farmasi Klinik Indonesia WMJ (Warmadewa Medical Journal) Global Medical and Health Communication Indonesian Journal of Science and Technology Jurnal Kebidanan The Indonesian Biomedical Journal IDJP (Indonesian Journal of Pharmaceutics) Jurnal Kedokteran Gigi Universitas Padjadjaran Universa Medicina Jurnal Bioteknologi & Biosains Indonesia (JBBI) Journal of Medicine and Health Molecular and Cellular Biomedical Sciences (MCBS) JURNAL PENDIDIKAN JASMANI DAN OLAHRAGA Syifa' MEDIKA: Jurnal Kedokteran dan Kesehatan Pharmacology and Clinical Pharmacy Research PengabdianMu: Jurnal Ilmiah Pengabdian kepada Masyarakat Jurnal Informatika Kaputama (JIK) Jurnal Ners Jurnal Ilmiah Farmako Bahari Jurnal Ilmu Faal Olahraga Indonesia Narra J Jurnal Pengabdian Olahraga Masyarakat JKKI : Jurnal Kedokteran dan Kesehatan Indonesia eJournal Kedokteran Indonesia Molekul: Jurnal Ilmiah Kimia Journal of Tropical Pharmacy and Chemistry Journal of Medicine and Health
Ronny Lesmana
Department Of Anatomy, Physiology And Biology Cell, Faculty Of Medicine, Universitas Padjadjaran, Sumedang 45363, Indonesia
Agriculture, Biological Sciences & Forestry Arts Humanities Biochemistry, Genetics & Molecular Biology Chemistry Computer Science & IT Control & Systems Engineering Dentistry Economics, Econometrics & Finance Education Engineering Environmental Science Health Professions Immunology & microbiology Languange, Linguistic, Communication & Media Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Nursing Public Health Social Sciences Veterinary Other

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Journal : Jurnal Bioteknologi

 1 
OPTIMASI TEKNIK WESTERN BLOT UNTUK DETEKSI EKSPRESI PROTEIN TANAMAN PADI (Oryza sativa L.) Susianti, .; Sukmana, Edi; Lesmana, Ronny; Supratman, Unang
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 2 (2019): December 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (350.722 KB) | DOI: 10.29122/jbbi.v6i2.3249

Abstract

Optimization of Western Blot Technique for Protein Expression of Rice Plant (Oryza sativa L.) Western blot (WB) technique has been widely used for analyzing protein expression and for identifying specific proteins derived from animals, plants, and microorganisms. During the use of WB, especially in agricultural studies, some difficulties are encountered such as unclear or unspecific protein bands, the presence of bubbles, and the absence of protein bands on membrane. This study aims to determine the WB conditions appropriate for the protein expression of rice plants (Oryza sativa L.). Protein from rice plant was extracted and the obtained protein lysate was then used for proteomic analysis using western blot with β-actin antibody. Our experiment showed that some optimized parameters like blocking buffers, the concentration of primary antibody and the ratio of secondary antibody determined the clarity of the results. β-actin was used as internal control that measured the success of the WB technique. Results showed that lysis process was important in determining good WB results in addition to the optimal blocking solution using a BSA of 0.2%, a primary antibody concentration of 1 μg mL–1, and a secondary antibody of 1:10,000. Optimizing techniques during extraction, incubation, and documentation facilitated good WB results.Keywords: β-actin; optimization; protein; rice plant; western blotABSTRAKTeknik western blot (WB) telah banyak digunakan untuk analisis ekspresi protein dan mengidentifikasi protein spesifik dari hewan, tumbuhan dan mikroorganisme. Dalam implementasi teknik WB, khususnya studi dalam bidang pertanian, beberapa kesulitan ditemui seperti pita protein tidak jelas, tidak spesifik, adanya gelembung, hingga tidak munculnya pita protein pada membran. Penelitian ini bertujuan untuk mengetahui kondisi WB yang tepat untuk deteksi protein tanaman padi (Oryza sativa L.). Protein tanaman padi diekstraksi, kemudian lysate protein yang didapat dianalisis dengan metode westernblot menggunakan antibody β-actin. Penelitian kami menunjukkan bahwa beberapa parameter yang dioptimasi seperti larutan blocking, konsentrasi antibodi primer dan rasio antibodi sekunder akan menentukan hasil yang jelas. β-actin digunakan sebagai kontrol internal yang menjadi tolok ukur keberhasilan teknik WB. Hasil menunjukkan bahwa proses lisis menjadi hal penting dalam menentukan hasil WB yang baik disamping larutan blocking yang optimal menggunakan BSA 0,2%, konsentrasi antibodi primer 1 µg mL–1 dan antibodi sekunder 1:10.000. Mengoptimalkan teknik selama ekstraksi, inkubasi dan dokumentasi membantu mendapatkan hasil WB yang baik.
OPTIMASI TEKNIK WESTERN BLOT UNTUK DETEKSI EKSPRESI PROTEIN TANAMAN PADI (Oryza sativa L.) . Susianti; Edi Sukmana; Ronny Lesmana; Unang Supratman
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 2 (2019): December 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (350.722 KB) | DOI: 10.29122/jbbi.v6i2.3249

Abstract

Optimization of Western Blot Technique for Protein Expression of Rice Plant (Oryza sativa L.) Western blot (WB) technique has been widely used for analyzing protein expression and for identifying specific proteins derived from animals, plants, and microorganisms. During the use of WB, especially in agricultural studies, some difficulties are encountered such as unclear or unspecific protein bands, the presence of bubbles, and the absence of protein bands on membrane. This study aims to determine the WB conditions appropriate for the protein expression of rice plants (Oryza sativa L.). Protein from rice plant was extracted and the obtained protein lysate was then used for proteomic analysis using western blot with β-actin antibody. Our experiment showed that some optimized parameters like blocking buffers, the concentration of primary antibody and the ratio of secondary antibody determined the clarity of the results. β-actin was used as internal control that measured the success of the WB technique. Results showed that lysis process was important in determining good WB results in addition to the optimal blocking solution using a BSA of 0.2%, a primary antibody concentration of 1 μg mL–1, and a secondary antibody of 1:10,000. Optimizing techniques during extraction, incubation, and documentation facilitated good WB results.Keywords: β-actin; optimization; protein; rice plant; western blotABSTRAKTeknik western blot (WB) telah banyak digunakan untuk analisis ekspresi protein dan mengidentifikasi protein spesifik dari hewan, tumbuhan dan mikroorganisme. Dalam implementasi teknik WB, khususnya studi dalam bidang pertanian, beberapa kesulitan ditemui seperti pita protein tidak jelas, tidak spesifik, adanya gelembung, hingga tidak munculnya pita protein pada membran. Penelitian ini bertujuan untuk mengetahui kondisi WB yang tepat untuk deteksi protein tanaman padi (Oryza sativa L.). Protein tanaman padi diekstraksi, kemudian lysate protein yang didapat dianalisis dengan metode westernblot menggunakan antibody β-actin. Penelitian kami menunjukkan bahwa beberapa parameter yang dioptimasi seperti larutan blocking, konsentrasi antibodi primer dan rasio antibodi sekunder akan menentukan hasil yang jelas. β-actin digunakan sebagai kontrol internal yang menjadi tolok ukur keberhasilan teknik WB. Hasil menunjukkan bahwa proses lisis menjadi hal penting dalam menentukan hasil WB yang baik disamping larutan blocking yang optimal menggunakan BSA 0,2%, konsentrasi antibodi primer 1 µg mL–1 dan antibodi sekunder 1:10.000. Mengoptimalkan teknik selama ekstraksi, inkubasi dan dokumentasi membantu mendapatkan hasil WB yang baik.
Co-Authors . SETIAWAN . Susianti Abd. Rasyid Syamsuri Abdul Hadi Hassan Achadiyani Agus Muji Santoso Agus Syahrani Agus Syahrani Ahmad Kurniawan Ahmad Kurniawan Ahmad Ramdan Ajeng Diantini Aldo Hartono Aldo, Mochammad Alia Bihrajihant Raya Alia Bihrajihant Raya Ambrosius Purba Ambrosius Purba Anang Wahid M.Diah Anas Subarnas Anas Subarnas Andrew Citrawan Anggraeni Dyah S Anggraeni Dyah Sulistiowati Anissa Anissa Anissa, Anissa Annisa F. Dewi Arief Budiono Arief Cahyanto Arief S. Kartasasmita Arifin Dwi Saputro Arto Yuwono Soeroto Astrid Feinisa Khairani Astrid, Stephanie Aulina, Datin Zahira Aziis Mardanarian Rosdianto Aziiz Mardanarian Rosdianto Aziiz Mardanarian Rosdianto Aziiz Rosdianto Azis Mardanarian Azmi, Mohamad Nurul Bachtiar, Willy Bahjah, Firda Hanan Baniasih, Akbar I. Baniasih, Akbar I. Bashari, Muhammad H. Bela Ita Karina Daba, Mariel Dani Sujana Darmayanti, Meita Decky Gunawan, Decky Dewi, Annisa F. Diana K Jasaputra Diana Krisanti Jasaputra Diana Krisanti Jasaputra Diandra Syifa Dimpuulina Erna Mariati Dwi Putri Larasati Edi Sukmana Edi Sukmana Edward J Hadi Endang Sutedja Endang Sutedja Enny Rohmawaty Euis Julaeha Evi Rinata Fajar Ari Widyatmoko Fani Rahma Yenita Fathurachman Fathurachman Fatona Suraya, Fatona Febriana Kurniasari Ferdyan Efza Fernandez, Veronica Fifi Veronica Filza Yulina Ade Finda Meyditia Finda Meyditia Fitri Fadhilah Galuga Sinalusur Sari Genta Syaifrin Laudza Gilang Muhamad Nur Iqbal Goenadi, Julia Windi Gunadi, Julia Windi Gunawan, Hanna Hadi Hadi Hana Goenawan Hanna Goeanawan Hanna Goenawan Hanna Goenawan Hanna Goenawan Hanna Gunawan Hanom Husni Syam Hendra Gunawan Hendy Setiawan Hendy Setiawan Herlina, Lien Herry Herman Hilmi Al Wahid Huda, Fathul Huda, Fathul I Made Joni Ida Ayu Putu Sri Widnyani Ieva B. Akbar Ieva Baniasih Akbar Ika M. Silviana Imam Megantara Indahwati, Dwiwahju Dian Irma Erika Herawati Irma Erika Herawati Irma Melyani Puspitasari Irma Ruslina Defi Isa Mahendra Ivan Christian Channel Ivanna Valentina Iwan Setiawan Iwan Setiawan Iwan Setiawan Iwan Setiawan Jatnika, Danti Dja Jihan Mudrika Rahmi Julia W Gunadi Julia Windi Gunadi Julia Windi Gunadi Jutti Levita Kanakarajah, Sarmmila Kartika Hajar Kirana, Kartika Hajar Kastella, Faysal Keiichi Motoyama Khairinisa, Miski A. Krisnadi, Sofie Riyani Kusmawan, Steven Zerin Putra Lazuardhi Dwipa Leonardo Lubis M.T., Vita Madhani Pradiptha Nugroho Maharani, Julia Maimun Syukri, Maimun Mariel Daba Mas Rizky A.A. Syamsunarno MEGANTARA, SANDRA Meita Dhamayanti Muchtaridi Muchtaridi Mulyanaga, William Giovanni Mutakin Mutakin Mutiarahmi, Citra Nur Nabilah Muhamad Naini, Al Arofatus Nanang Wahyudin Nanang Wahyudin Nanny N. N. M. Soetedjo Nasrul Wathoni Norisca Aliza Putri Nova Sylviana Nova Sylviana Nuraini Yasmin Nuroh Najmi Nurul Fadhilah Oki Suwarsa Okta Wismandanu Oktorina, Lenny Pangestuti, Balqist Sriprobo Pardede, Akim Manaor Hara Penny Setyawati Martioso Permadi, Andieni Faqhira Permana, Noah Jefferson Permana, Shenny S.S Pinkan Al Shabrina Pitu Wulandari Prasetyo, Wibowo Budi Prayudi Santoso Putri Tessa Putri, Aufa A.D Qiantori, Adziqa Ammara Radhiyanti, Putri Teesa Rahdiyanti, Putri Teesa Ramadhanti, Julia Ramdani Hamzah Ray, Hamidie Ronald Daniel Rejeki, Purwo Sri Renaldi Prasetia Reni Farenia Reni Farenia rima siburian Riska A Safira Risna Agustina Risna Agustina Roro Wahyudianingsih Rosdianto, Aziis Mardanarian Rosdianto, Aziiz M. Rosdianto, Aziiz Mardanarian Rusdin, Agus Ruswana Anwar Sandra Amalia Riyadi Santi Sofiyanti Sarmmila Kanakarajah Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Shelly Iskandar Silviana, Ika M. Simanungkalit, Jayne Mary Siti Baitul Mukarromah Siti Mukarromah Siti Nuriyatus Zahrah Slamet Imron Sri Adi Sumiwi Steffi Liem Sudigdo Adi Sudrajat, Ratu Z.A.P sugianli, Adhi Sukmayani, Jasmine Y. Sunarjati Soedigdoadi Sunarjati Sudigdo Adi Sunarjati Sudigdo Adi Sunarjati Sudigdoadi Sunaryati S. Adi Supriatno Salam Susanti Husen Susianti Susianti Susianti Susianti Susiarno, Hadi Tarawan, Vita M. Tarawan, Vita M. Teresa Lucretia, Teresa Teti Madiadipoera TIANA MILANDA Tina Rostinawati Titing Nurhayati Tjahjono D. Gondhowiardjo Tri Mayanti Trianing Tyas Kusuma Tyagita Hartady Unang Supratman Vita M. Tarawan Vita Murniati Tarawan Vita Murniati Tarawan Vita Murniati Tarawan Lubis Vita Mutia Tarawan Wahyudianigsih, Roro Wahyuni, Indah S. Willy Bachtiar Yanuar Dwi Prastyo Yasmin, Nuraini Yenny Noor Yudi Feriandi Yulia Sofiatin Yumilia, Hoo Yunas, Aisya Astri Yuni Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi, Yuni Susanti Yusuf, Muchamad R.A Zaini, Lia M. Zakiyah, Neily Zulhendri, Felix