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Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan

Proteomic study related with placental protein has been an interest topic to the researcher over decades to determine the function and mechanism of particular protein that perhaps to improve pregnancy related disorder or pregnancy diagnosis in human and livestock industry. Protein extraction is a pr...

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Main Author: Tee Yung Yee
Format: Undergraduate Final Project Report
Language:English
Published: 2019
Online Access:http://discol.umk.edu.my/id/eprint/4848/1/TEE%20YUNG%20YEE.pdf
http://discol.umk.edu.my/id/eprint/4848/
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spelling my.umk.eprints.48482022-05-27T03:05:44Z http://discol.umk.edu.my/id/eprint/4848/ Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan Tee Yung Yee Proteomic study related with placental protein has been an interest topic to the researcher over decades to determine the function and mechanism of particular protein that perhaps to improve pregnancy related disorder or pregnancy diagnosis in human and livestock industry. Protein extraction is a preliminary step of protein purification which mainly focus on maximisation of total protein yield. The heterogenous properties causes diversification of protein, therefore there is no absolute protocol in protein extraction, the type of buffer and ratio could give different protein concentration in different type of mammalian tissue, hence lead to the study of optimisation of types of buffer and buffer ratio to obtain better total protein yield. The objectives of this study are to compare the total protein yield based on three types of buffer with three different ratios. Three types of buffer included phosphate buffer saline (PBS), radioimmunoprecipitation assay (RIPA) buffer and RIPA buffer with addition of protease inhibitor (Pi) whilst sample to buffer ratio, 1:1, 1:3, and 1:5 were used in the series of protein extraction process that involved mechanical disruption, incubation, sonication, and centrifμgation. Bradford assay was carried out to determine the total protein yield based on the standard curve of bovine serum albumin (BSA). From this study it is concluded there is a significant interaction between buffer type and ratio (P<0.05) where the use RIPA buffer with 1:1 ratio gave the best total protein yield (194.880 ± 15.089 mg/g). This study is perhaps to be a useful reference to future study in term of protein extraction of mammalian tissue especially placental tissue. 2019 Undergraduate Final Project Report NonPeerReviewed text en http://discol.umk.edu.my/id/eprint/4848/1/TEE%20YUNG%20YEE.pdf Tee Yung Yee (2019) Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan. Final Year Project thesis, University Malaysia Kelantan. (Submitted)
institution Universiti Malaysia Kelantan
building Perpustakaan Universiti Malaysia Kelantan
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Kelantan
content_source UMK Institutional Repository
url_provider http://umkeprints.umk.edu.my/
language English
description Proteomic study related with placental protein has been an interest topic to the researcher over decades to determine the function and mechanism of particular protein that perhaps to improve pregnancy related disorder or pregnancy diagnosis in human and livestock industry. Protein extraction is a preliminary step of protein purification which mainly focus on maximisation of total protein yield. The heterogenous properties causes diversification of protein, therefore there is no absolute protocol in protein extraction, the type of buffer and ratio could give different protein concentration in different type of mammalian tissue, hence lead to the study of optimisation of types of buffer and buffer ratio to obtain better total protein yield. The objectives of this study are to compare the total protein yield based on three types of buffer with three different ratios. Three types of buffer included phosphate buffer saline (PBS), radioimmunoprecipitation assay (RIPA) buffer and RIPA buffer with addition of protease inhibitor (Pi) whilst sample to buffer ratio, 1:1, 1:3, and 1:5 were used in the series of protein extraction process that involved mechanical disruption, incubation, sonication, and centrifμgation. Bradford assay was carried out to determine the total protein yield based on the standard curve of bovine serum albumin (BSA). From this study it is concluded there is a significant interaction between buffer type and ratio (P<0.05) where the use RIPA buffer with 1:1 ratio gave the best total protein yield (194.880 ± 15.089 mg/g). This study is perhaps to be a useful reference to future study in term of protein extraction of mammalian tissue especially placental tissue.
format Undergraduate Final Project Report
author Tee Yung Yee
spellingShingle Tee Yung Yee
Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
author_facet Tee Yung Yee
author_sort Tee Yung Yee
title Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
title_short Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
title_full Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
title_fullStr Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
title_full_unstemmed Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan
title_sort optimisation of total protein concentration based on buffer type and ratio in placental protein extraction of bovine kedah kelantan
publishDate 2019
url http://discol.umk.edu.my/id/eprint/4848/1/TEE%20YUNG%20YEE.pdf
http://discol.umk.edu.my/id/eprint/4848/
_version_ 1763303471456452608
score 13.211869

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