Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review
Advancements in technology and analytical methods enable researchers to explore the biochemical events that cause variation in meat quality. Among those, western blot techniques have been successfully used in identifying and quantifying the key proteins that have critical functions in the developmen...
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| Format: | Article |
| Language: | English |
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Iowa State University Digital Press
2022-02-01
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| Series: | Meat and Muscle Biology |
| Subjects: | |
| Online Access: | https://www.iastatedigitalpress.com/mmb/article/id/11551/ |
| _version_ | 1797222188546260992 |
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| author | Chaoyu Zhai Elisabeth J. Huff-Lonergan Mahesh N. Nair Steven M. Lonergan |
| author_facet | Chaoyu Zhai Elisabeth J. Huff-Lonergan Mahesh N. Nair Steven M. Lonergan |
| author_sort | Chaoyu Zhai |
| collection | DOAJ |
| description | Advancements in technology and analytical methods enable researchers to explore the biochemical events that cause variation in meat quality. Among those, western blot techniques have been successfully used in identifying and quantifying the key proteins that have critical functions in the development of meat quality. Housekeeping proteins, like β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and tubulins are often used as internal controls in western blots to normalize the abundance of the protein of interest. However, there are increasing concerns about using housekeeping proteins for western blot normalization, as these proteins do not demonstrate any loading differences above the relatively small total protein loading amounts of 10μg. In addition, the interaction between these housekeeping proteins and programmed cell death processes highlights the concerns about using the housekeeping protein as the internal control in meat quality research. Moreover, recent proteomic research has indicated that the abundance of some housekeeping proteins, like β-actin, GAPDH, and tubulin, can be altered by preslaughter stress, dietary supplementation, sex, slaughter method, genotype, breed, aging period, muscle type, and muscle portion. Furthermore, these housekeeping proteins could have differential expression in meat with differing color stability, tenderness, and water holding capacity. Therefore, this review aims to examine the realities of using housekeeping proteins as the loading control in meat quality research and introduce some alternative methods that can be used for western blot normalization. |
| first_indexed | 2024-04-24T13:17:22Z |
| format | Article |
| id | doaj.art-94a2c1fb005f435cbd7393d5e68d135b |
| institution | Directory Open Access Journal |
| issn | 2575-985X |
| language | English |
| last_indexed | 2024-04-24T13:17:22Z |
| publishDate | 2022-02-01 |
| publisher | Iowa State University Digital Press |
| record_format | Article |
| series | Meat and Muscle Biology |
| spelling | doaj.art-94a2c1fb005f435cbd7393d5e68d135b2024-04-04T17:20:59ZengIowa State University Digital PressMeat and Muscle Biology2575-985X2022-02-016110.22175/mmb.11551Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini ReviewChaoyu Zhai0https://orcid.org/0000-0001-5148-4584Elisabeth J. Huff-Lonergan1https://orcid.org/0000-0001-5012-3277Mahesh N. Nair2https://orcid.org/0000-0002-2917-1028Steven M. Lonergan3Colorado State UniversityDepartment of Animal Science, Iowa State UniversityDepartment of Animal Sciences, Colorado State UniversityIowa State UniversityAdvancements in technology and analytical methods enable researchers to explore the biochemical events that cause variation in meat quality. Among those, western blot techniques have been successfully used in identifying and quantifying the key proteins that have critical functions in the development of meat quality. Housekeeping proteins, like β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and tubulins are often used as internal controls in western blots to normalize the abundance of the protein of interest. However, there are increasing concerns about using housekeeping proteins for western blot normalization, as these proteins do not demonstrate any loading differences above the relatively small total protein loading amounts of 10μg. In addition, the interaction between these housekeeping proteins and programmed cell death processes highlights the concerns about using the housekeeping protein as the internal control in meat quality research. Moreover, recent proteomic research has indicated that the abundance of some housekeeping proteins, like β-actin, GAPDH, and tubulin, can be altered by preslaughter stress, dietary supplementation, sex, slaughter method, genotype, breed, aging period, muscle type, and muscle portion. Furthermore, these housekeeping proteins could have differential expression in meat with differing color stability, tenderness, and water holding capacity. Therefore, this review aims to examine the realities of using housekeeping proteins as the loading control in meat quality research and introduce some alternative methods that can be used for western blot normalization.https://www.iastatedigitalpress.com/mmb/article/id/11551/postmortem muscleβ-actinGAPDHtubulinwestern blot |
| spellingShingle | Chaoyu Zhai Elisabeth J. Huff-Lonergan Mahesh N. Nair Steven M. Lonergan Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review Meat and Muscle Biology postmortem muscle β-actin GAPDH tubulin western blot |
| title | Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review |
| title_full | Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review |
| title_fullStr | Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review |
| title_full_unstemmed | Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review |
| title_short | Housekeeping Proteins in Meat Quality Research: Are They Reliable Markers for Internal Controls in Western Blot? A Mini Review |
| title_sort | housekeeping proteins in meat quality research are they reliable markers for internal controls in western blot a mini review |
| topic | postmortem muscle β-actin GAPDH tubulin western blot |
| url | https://www.iastatedigitalpress.com/mmb/article/id/11551/ |
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