Western Blot

Year 2021, Volume: 6 Issue: 1, 45 - 56, 30.04.2021

Hatice Pelin Aslım , Oya Bulut

https://doi.org/10.31797/vetbio.799660

Abstract

The Western Blot technique is an old and reliable method used in molecular biology to uniquely detect a single protein from any sample. It is a frequently used method for investigating the responses of proteins to stimuli, evaluating protein expression as either absent or increased-decreased, determining cell specific protein isoforms and determining proteins responsible for the occurrence of genetic diseases. In this composition, , information has been provided about principle, protocol and use of Western Blot technique in virology.

Keywords

Method, Virology, Western Blot

Project Number

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Western Blot

Abstract

Western Blot tekniği, herhangi bir örnekten istenen tek bir proteini özgün bir şekilde saptamak için moleküler biyolojide kullanılan eski ve güvenilir bir yöntemdir. Proteinlerin uyaranlara karşı verdiği yanıtların araştırılmasında, protein ifadesinin var-yok ya da artmış-azalmış olarak değerlendirilmesinde, hücre spesifik protein izoformlarının belirlenmesinde, genetik hastalıkların ortaya çıkışından sorumlu proteinlerin belirlenmesinde sıklıkla kullanılan bir metottur. Yapılan bu derlemede, Western Blot tekniğinin prensibi, protokolü ve virolojide kullanımı hakkında bilgi verilmiştir.

Keywords

Metot, Violoji, Western Blot

Supporting Institution

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Project Number

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Thanks

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References

• Anonim, (2009). Polyacrylamide gel electrophoresis. https://en.wikipedia.org/wiki/Polyacrylamide gel electrophoresis Erişim tarihi: 27.07.2020
• Anonim, (2020a, 27 Temmuz). Western blot. https://en.wikipedia.org/wiki/Western_blot#:~:text=SDS%20is%20generally%20used%20as,%2C%20negatively%2C%20or%20neutrally%20charged.&text=The%20gel%20electrophoresis%20step%20is,the%20cross%2Dreactivity%20of%20antibodies
• Anonim, (2020b, 27 Temmuz). Choosing a primary antibody or secondary antibody. https://www.abcam.com/protocols/choosing-an-antibody
• CUSABIO, (2018, 27 Temmuz 2020 ). Laboratory technical manual western blotting. Erişim adresi: https://www.cusabio.com/protocols/Western_Blotting(WB)_Protocol.pdf
• Flores, M.K., Salazar-Anzures, Á.T., Fernández-Torres, J., Pineda, C., Aguilar-González, C.A., López-Reyes, A. (2017). Western blot: a tool in the biomedical field. Investigacion en Discapacidad. 3, 128-137.
• Hassan, A.O., Vemula, S.V., Sharmal, A., Bangaril, D.S., Mishral, K.K., Mitta, S.K. (2017). 155R is a novel structural protein of bovine adenovirus type 3, but it is not essential for virus replication. Journal of General Virology, 98, 749-753.
• He, Q., Chong, K.H., Chng, H.K., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E.İ., Kwang, J. (2004). Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. American society for mıcrobıyolog. 11, 417-422.
• Hemmatzadeh, F., Kazemimanesh, M. (2017). Detection of specific antigens of newcastle disease virus using an absorbed Western blotting method. Iranian Journal of Veterinary Research, 18, 92-96.
• Jayme, S.I., Field, H.E., Jong, C.D., Olival, K.J., Marsh, G., Tagtag, A.M., Hughes, T., Bucad, A.C., Barr, J., Azul, R.R., Retes, L.M., Foord, A., Yu, M., Cruz, M.S., Santos, I.J., Lim, T.M.S., Benigno, C.C., Epstein, J.H., Wang, L.F., Daszak, P., Newman, S.H. (2015). Molecular evidence of Ebola Reston virus infection in Philippine bats. Virology Journal,, 12,107.
• Jensen, E.C. (2012). The Basics of Western Blotting. The Anatomical Record, 295, 369–371.
• Kennedy, M. (2005). Methodology in diagnostic virology. Veterinary Clinics: Exotic Animal Practice, 8, 7–26.
• Goldberg, T.L., Chapman, C.A., Cameron, K., Saj, T., Karesh, W.B., Wolfe, N.D., Wong, S.W., Dubois, M.E., Slifka, M. (2008). Serologic evidence for novel poxvirus in endangered red colobus monkeys, Western Uganda. Emerging Infectious Disease,. 5, 801.
• Kurien, B.T., Scofield, R. (2006). Western blotting. Methods, 38, 283-293.
• Smith A.G., Jayaram J, Johnson C.B., Ellis E.A., Vitha S., Collisson E.W., Holzenburg A. (2009). Improved Protein Detection Using Cold Microwave Technology. In: Protein blotting and detection methods and protocols. Ed. Kurien, B.T., Scofield, R. (pp. 533-544). Human Press, Oklohama City, OK, USA.
• Li, G., Kawashima, H., Ogose, A., Ariizumi, T., Xu, Y., Hotta, T., Urata, Y., Fujiwara, T., Endo, N. (2011). Efficient virotherapy for osteosarcoma by telomerase-specific oncolytic adenovirus. Journal of Cancer Research and Clinical Oncology, 137, 1037-1051.
• Liu, Q., Suresh, K.T., Babiuk, L.A. (2001). Nuclear Localization of the ORF2 Protein Encoded by Porcine Circovirus Type 2. Virology, 285, 91–99.
• MacPhee, D.J. (2010). Methodological considerations for improving Western blot analysis. Journal of Pharmacological and Toxicological Methods, 61, 171-177.
• Mahmood, T., Yang, P.C. (2012). Western Blot: Technique, Theory, and Trouble Shooting. North American Journal Medical Sciences. 9, 429–434.
• Martínez-Flores, K., Salazar-Anzures, Á.T., Fernández-Torres, J., Pineda, C., Aguilar-González, C.A., López-Reyes, A. (2017). Western blot: a tool in the biomedical field. Investigacion en Discapacidad, 3, 128-137.
• Martínez-Maqueda, D., Hernández-Ledesma, B., Amigo, L., Miralles, B., Gómez-Ruiz, J.Á. (2013). Extraction/fractionation techniques for proteins and peptides and protein digestion. In: Proteomics in Foods. Ed: Toldra F., Nollet L.M.L., (pp. 21-50). Springer, New York, USA.
• Martins, T.B., Woolstenhulme, R.D., Jaskowski, T.D., Hill, H.R., Litwin, C.M. (2001). Comparison of four enzyme immunoassays with a Western blot assay for the determination of type-specific antibodies to herpes simplex virus. Microbiology and Infectious Disease, 2, 272-277.
• Melo, D.M., Payant, B., Liu, W.C., Uhl, S., Hoagland, D., Moller, R., Jordan, T.X., Oishi, K., Panis, M., Sachs, D., Wang, T.T., Schwartz, R.E., Lim, J.K., Albrecht, R.A., TenOever, B.R. (2020). Imbalanced Host Response to SARS-CoV-2 Drives Development of COVID-19. Cell, 181, 5, 1036-1045.
• Memelink, J., Swords, K.M.M., Staehelın, L.A., Hoge, J.H.C. (1994). Southern, Northern and Western blot analysis. Plant Molecular Biology Manual, l, 1-23.
• Minafra, A., Casati, P., Elicio, V., Rowhani, A., Saldarelli, P., Savino, V., Martelli, G.P. (2000). Serological detection of Grapevine rupestris stem pitting-associated virus (GRSPa V) by a polyclonal antiserum to recombinant virus coat protein. Vitis, 3, 115-118.
• Sapan, C.V., Lundblad, R.L., Price, N.C. (1999). Colorimetric protein assay techniques. Biotechnology and Applied Biochemistry. 29, 99-108.
• Schaller, O., Fatzer, R., Stack, M., Clark, J., Cooley, W., Biffiger, K., Egli, S., Doherr, M., Vandevelde, M., Heim, D., Oesch, B., Moser, M. (1999). Validation of a Western immunoblotting procedure for bovine PrPSc detection and its use as a rapid surveillance method for the diagnosis of bovine spongiform encephalopathy (BSE). Acta Neuropathologica, 98, 437–443.
• Schägger H. (2006). Tricine–sds-page. Nature Protocols. 1, 1-16.
• Siedler, B.S., Roloff, B.C., Sa, G.L.D., Neis, A., Conceiçao, F.R., Hartwig, D.D., Borsuk, S., Dellagostin, O.A., Campos, F.S., Roehe, P. M., Hartleben, C.P., McBride, A.J.A. (2017). Secretory expression of bovine herpesvirus type 1/5 glycoprotein E in Pichia pastorisfor the differential diagnosis of vaccinated or infected cattle. Protein Expression and Purification. 130, 21-27.
• Schlickeiser, S., Pleyer, U. (2007). Western, Northern, and Southern Blotting. In: Basic Science Techniques in Clinical Practice. Ed: Patel, H.R.H., Arya, M., Shergill, I.S. (pp. 48-57). Springer, London, England.
• Sosnovtsev, S.V., Green, K.Y. (2000). Identification and Genomic Mapping of the ORF3 and VPg Proteins in Feline Calicivirus Virions. Virology, 277, 193–203.
• Streckert, H.J., Bürüssow, H., Sure, K., Werchau, H. (1986). Antipeptide Antibodies Directed Against the Carboxy-Terminal Region of SV40 Structural Proteins VP2 and VP3. Journal of Cellular Biochemistry, 31, 277-287.
• Thiry, D., Mauroy, A., Saegerman, C., Licoppe, A., Fett, T., Thomas, I., Brochier, B., Thiry, E., Linden, A. (2017). Belgian Wildlife as Potential Zoonotic Reservoir of Hepatitis E Virus. Transboundary and Emerging Diseases, 64, 764–773.
• Whetstone, C.A., VanDerMaaten, M.J., Miller, J.M. (1991). A western blot assay for the detection of antibodies to bovine immunodeficiency-like virus in experimentally inoculated cattle, sheep, and goats. Archives of Virology, 116, 119-131.
• Woo, P.C.Y., Lau, S.K.P., Tsoi, H-w., Chan, K-h., Wong, B.H., Che, X-y., Tam, V.K.P., Tam, S.C.F., Cheng, V.C.C., Hung, I.F.N. (2004). Relative rates of non-pneumonic SARS coronavirus infection and SARS coronavirus pneumonia. The Lancet, 363, 841-845.