Abstract
Western blotting is used to transfer proteins from polyacrylamide gels (PAGs) to the membranes, making its identification and determination more convenient. For this, PAG and membrane are sandwiched together between two parallel electrodes. On passage of current, proteins transfer from the gels to the membranes. This chapter outlines various components, merits and demerits of western blotting technique. Most commonly used blotting membranes, namely, nitrocellulose membranes, polyvinylidene difluoride (PVDF), and nylon, are also compared for their properties, merits and demerits in this section. Further, visualization and detection of protein markers and targeted proteins over the membrane can be achieved using various protocols. Ponceau S or Congo red dyes are frequently employed for the detection of protein markers, whereas for the detection of analyte (specific protein) antigen antibody interaction, enzyme-based interaction of carbohydrate moiety reaction can be utilized. Lastly, its applications in dairy science are also outlined.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
https://info.gbiosciences.com/blog/how-to-prepare-samples-for-western-blot-analysis-1
Healthcare GE (2011) Western blotting: principles and methods. In: Handbooks from GE Healthcare. Sweden
Kurien BT, Scofield RH (2015) Western blotting: methods and protocols. Springer, Heidelberg
Reinhardt TA et al (2013) Bovine milk proteome: quantitative changes in normal milk exosomes, milk fat globule membranes and whey proteomes resulting from Staphylococcus aureus mastitis. J Proteome 82:141–154
El-Agamy EI et al (2009) Are camel milk proteins convenient to the nutrition of cow milk allergic children? Small Rumin Res 82(1):1–6
Chávez NA et al (2008) Detection of bovine milk adulterated with cheese whey by western blot immunoassay. Food Agric Immunol 19(4):265–272
Neelima S, Rajan Y, Rajput S, Mann B (2013) Chemical and functional properties of glycomacropeptide (GMP) and its role in the detection of cheese whey adulteration in milk: a review. Dairy Sci Technol 93(1):21–43
Author information
Authors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Gandhi, K., Sharma, N., Gautam, P.B., Sharma, R., Mann, B., Pandey, V. (2022). Western Blotting. In: Advanced Analytical Techniques in Dairy Chemistry. Springer Protocols Handbooks. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1940-7_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1940-7_5
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-0716-1939-1
Online ISBN: 978-1-0716-1940-7
eBook Packages: Springer Protocols