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All Journal HAYATI Journal of Biosciences Jurnal Teknologi Dan Industri Pangan VALENSI Jurnal Kimia (Journal of Chemistry) Jurnal Veteriner JURNAL KIMIA SAINS DAN APLIKASI Biotika: Jurnal Ilmiah Biologi Bionatura Indonesian Journal of Applied Sciences Jurnal Ilmu Dasar Jurnal Natur Indonesia Majalah Kedokteran Bandung Cropsaver : Journal of Plant Protection Jurnal Penelitian Hasil Hutan Indonesian Journal of Cancer Chemoprevention Journal of Mathematical and Fundamental Sciences Indonesian Journal of Pharmaceutical Science and Technology STOMATOGNATIC- Jurnal Kedokteran Gigi Indonesian Journal of Science and Technology Dentino: Jurnal Kedokteran Gigi The Indonesian Biomedical Journal Padjadjaran Journal of Dentistry Jurnal Bioteknologi & Biosains Indonesia (JBBI) JURNAL PENDIDIKAN JASMANI DAN OLAHRAGA Al-Kimia Chimica et Natura Acta ALCHEMY Jurnal Penelitian Kimia Indonesian Journal of Chemistry Communications in Science and Technology Akta Kimia Indonesia Jurnal Kartika Kimia B-Dent, Jurnal Kedokteran Gigi Universitas Baiturrahmah Molekul: Jurnal Ilmiah Kimia Jurnal Ilmu dan Teknologi Kayu Tropis Jurnal Permukiman Jurnal Penelitian Hasil Hutan Jurnal Ilmiah Berkala: Sains dan Terapan Kimia Denta: Jurnal Kedokteran Gigi Makara Journal of Science Jurnal Ilmiah Sains Molekul: Jurnal Ilmiah Kimia Jurnal Penelitian Hasil Hutan
Unang Supratman
Departemen Kimia Fakultas Matematika Dan Ilmu Pengetahuan Alam Universitas Padjadjaran Central Laboratory Of Universitas Padjadjaran, Jatinangor 45363, Indonesia
Agriculture, Biological Sciences & Forestry Astronomy Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Dentistry Earth & Planetary Sciences Education Energy Engineering Environmental Science Health Professions Immunology & microbiology Industrial & Manufacturing Engineering Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Physics Public Health Veterinary

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Journal : Jurnal Bioteknologi

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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 , ,. Melanie - Ruchiyat . Horizon . Susianti Ace Tatang Hidayat Ace Tatang Hidayat Ace Tatang Hidayat Ace Tatang Hidayat Achmad Zainuddin Achmad Zainuddin Ade Akbar Abdilla Ade Kania Ningsih Ade Kania Ningsih, Ade Kania Adel Zamri Agus Safari Agus Safari Agus Safari Agus Safari Agustini, Dewi Meliati Ahmad Darmawan Ahmad Kurniawan Ahmad Kurniawan Ahmad Ramdan Ahmad Yani Nelly Wahyuni Lia Destiarti Akhmad Darmawan Al Arofatus Naini Al Arofatus Naini Al Arofatus Naini Aldo Hartono Almas Widyana Alya Tsamrotul Amir M. Suruwaky Anas Subarnas Anas Urbanas Anastasya Firdausi Andi Rahim Andre A. Sonda Aneu Wahyuni Anjari, Intan Hawina Aprilia Permata Sari Aprilia Permata Sari Aprina, Lutfia Silva Ari Hardianto Ari Hardianto Ari Widiyantoro Arif Rahman Hakim Arif Rahman Hakim Arif Rahman Hakim Arif Rahman Hakim Arlette Setiawan Arlette Setiawan Arlette Setiawan Arlette Setiawan Arlette Setiawan Arlette Suzy Puspa Pertiwi Arnov, Steffi Triany Arto Yuwono Soeroto Asep Supriadin Asri Peni Wulandari Asri Peni Wulandari, Asri Peni Astrid Feinisa Khairani Aziiz Mardanarian Rosdianto Azmi Azhari Azmi Azhari, Azmi Azmi, Mohamad Nurul Benny Joy Betry Pujiastuti Budiman, Yudha P. C Hanny Wijaya Celcius Waranmaselembun Christina Marpaung Dadan Sumiarsa Danar Dono Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati Darwati, Darwati, Darwati Deden Indra Dinata Deden Indra Dinata, Deden Indra Desi Harnet Desi Harneti Desi Harneti Desi Harneti Putri Huspa Desi Harneti Putri Huspa Dewa G Katja Dewa Gede Katja Dewa Gede Katja Dewa Gede Katja Dikdik Kurnia Dimpuulina Erna Mariati Dini Oktaviani Dondin Sajuthi Dudi Runadi DUDI RUNADI Edi Suanto, Edi Edi Sukmana Edi Sukmana Edi Sukmana Elis Hastuti Elisabeth Krismayanti Elvi Rusmiyanto Pancaning Wardoyo, Suci Lestari, Mukarlina, Erina Hilmayanti Erina Hilmayanti Erina Hilmayanti Erina Hilmayanti Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Euis Julaeha Evan Hadrian Ezatul Ezleen Kamarulzaman Fadlilah, Gina Faizah Maira Faizal Hermanto Fajar Fauzi Abdullah Fajar Fauzi Abdullah Fajar Fauzi Abdullah Fajar Fauzi Abdullah Fajar Fauzi Abdullah Fajar Fauzi Abdullah Fajar, Mohamad Fajar, Mohamad Fani Rahma Yenita Farabi, Kindi Fathurachman Fathurachman Fauzan Zein Muttaqien Ferdyan Efza Ferry Ferdiansyah Sofian, Ferry Ferdiansyah Filza Yulina Ade Fizrul Indra Lubis Galih Bayu Pratama Galuga Sinalusur Sari Ghina Izdihar Gilang Muhamad Nur Iqbal Gunawan, Latifah Hadi Kuncoro Hadi Kuncoro Hana Goenawan Hanna Goeanawan Hanna Goenawan Harizon Harizon Harlia Harlia Hasnah Osman Hasnah Osman Hedi Paramita Herdanu Rizqullah Hersa Milawati Hersa Milawati Hersa Milawati Hideo Hayashi Hideo Hayashi Hideo Hayashi Hideo Hayashi Hideo Hayashi Hideo Hayashi Hilmayanti, Erina Huda, Muhammad Badrul Husein Hernadi Bahti Hutagaol, Ricson Pemimpin Ichsan Nurul Bari Ida Ayu Putu Sri Widnyani Ida Nur Farida Ida Nur Farida Ida Nurfarida Ihsan Rahadian Iis Intan Widiyowati Indri Indriyani Indriyani, Indri Inne Suherna Sasmita Intan Hawina Anjari Intan Rahmayanti Iqbal Musthapa Iqbal Wahyu Mustaqim Isa Mahendra Isramiharti Isramiharti Iwan Setiawan Iwan Setiawan Iwan Setiawan Jamaludin Al-Anshori Jihan Mudrika Rahmi Julia Windi Gunadi Julia Windi Gunadi Julinton Sianturi Julinton Sianturi Kadarusman Kansy Haikal Kansy Haikal Kautsari, Arsi Khadijah Awang Khairunnisa, Shofiyah Khalijah Awang Khalijah Awang Khalijah Awang Khalijah Awang Khalijah Awang Khalijah Awang Khalijah Awang Khlaijah Awang Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kindi Farabi Kok Tong Wong Kristin Shinta Dewi Kusumiyati Laode Rijai Lilis Siti Aisyah Lilis Siti Aisyah Lilis Siti Aisyah Lilis Siti Aisyah, Lilis Siti Lindung Tri Puspasari Lindung Tri Puspasari M. S. Soedjanaatmadja Maira, Faizah Mantiri, Sisilia A Maryati Maryati Mas Rizky A.A. Syamsunarno Max R.J Runtuwene Max R.J Runtuwene Mayshah Purnamasari MEGANTARA, SANDRA Milawati, Hersa Moelyono Moektiwardoyo Mohamad Fajar Mohamad Fajar Mohamad Nurul Azmi Mohamad Nurul Azmi Mohamad Nurul Azmi Mohamad Nurul Azmi Mohamad Nurul Azmi Mohamad Nurul Azmi Mohamad Nurul Azmi bin Mohamad Taib Mohamad Nurul Azmi Mohamad Taib Mohamad Nurul Azmi Mohamad Taib Mohamed Ashraf Ali Mohd. Zaheen Hassan Mohd. Zaheen Hassan Muchlis, Handi Nugraha Muhamad Nurul Azmi Muhamad Nurul Azmi Muhammad Hanafi Muhammad Hanafi Muhammad Hanafi Muhammad Solehin Abd Ghani Muhammad Solehin Abd Ghani Murtihapsari . Mustaqim, Iqbal Wahyu Mustofa, Hidayat Nurul Muttaqin, Fauzan Zein Nadia Mohamed Yusoff Nadya Thufaila Nafiah, Mohamad Azlan Naini, Al Arofatus Nasrudin Nasrudin Nayla Haraswati Nayla Haraswati Noor Rain Abdullah Noor Rain Abdullah Nova Sylviana Nova Sylviana Nulelasari Nurlelasari Nunung Kurniasih Nunung Kurniasih Nunung Kurniasih, Nunung Nur Insani Amir Nur Muhammad Miftah Nurabi Ferdiana Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Nurlelasari Okta Wismandanu Paramita, Hedi Ponis Tarigan Prasetyo, Wibowo Budi Pratama, Galih Bayu Prayudi Santoso Primahana, Gian Purbaya, Sari Purnama Purnama Purnama Purnama Purnama Purnama Purwoko, Agus Puspita Sari Putri, Ghesta Alifka Rahmawati Rahmawati Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Rani Maharani Ph.D Ratu Safitri Ray, Hamidie Ronald Daniel Rejeki, Purwo Sri Revan Hardiawan Richa Mardianingrum Ricson Pemimpin Hutagaol Rika Meliansyah RIKA MELIANSYAH Risyandi Anwar Risyandi Anwar Riza Apriani Rizky Abdulah Rizky Abdullah Romundza, Febbry Ronauli Fitriana Ronny Lesmana Ronny Lesmana Roosje Rosita Oewen, Roosje Rosita Roro Wahyudianingsih Rudi Hendra Rymond Jusuf Rumampuk Safri Ishmayana Safriansyah, Wahyu Salam, Supriatno Samuel San Parulian Sandra Amalia Riyadi Sari Purbaya Sari Purbaya Sari, Aprilia Permata Satwika Nandiwardhana Setiawan Setiawan Setiawan Setiawan Setiawan Setiawan Shabarni Gaffar Shabarni Gaffar Shiono, Yoshihito Sianturi, Julinton Sinaga, Siska Elisahbet Sisilia A Mantiri Siska Elisahbet Sinaga Siska Mulya Octavia Siska Mulya Octavia Siti Hani Pratiwi Sofa Fajriah Sofa Fajriah Sofa Fajriah Sofa Fajriah Sofa Fajriah Sofa Fajriah Sofa Fajriah Srikandi, Srikandi Subekti Mauluddin Sudarjat Sudarjat Sunarjati Sudigdoadi Supriatno Supriatno Supriatno Salam Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriatno Supriyatna Supriyatna Supriyatna Sutardjo Supriyatna Sutardjo Supriyatna Sutardjo Supriyatna Sutardjo Supriyatna Sutardjo Supriyatna, - Suseno Amien Susianti Susianti Susianti Susianti Susianti Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Tati Herlina Teddy Budiyansyah Thaigarajan Parumasivam Tiara Prima Amalya Tjandrawati Mozef Toto Subroto Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Mayanti Tri Reksa Saputra Tri Reksa Saputra Tri Reksa Saputra, Tri Reksa Vicki Nishinarizki Vidia Afina Nuraini Vita Murniati Tarawan Vita Murniati Tarawan W.C. Taylor Wahyu Syafriansyah Wahyuni, Aneu Wawan Hermawan Wawan Hermawan Winda Sukmawati Winda Sukmawati Witriany Rayapratiwi Yeni Mulyani Yenny Febriani Yun Yenny Febriani Yun Yenny Febriani Yun Yenny Febriani Yun, Yenny Febriani Yeong Keng Yoon Yessi Permana Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yoshihito shiono Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yoshihito Shiono Yum Eryanti Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi Yuni Susanti Pratiwi, Yuni Susanti Yusup Hidayat