Staff View: Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot

Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot

Targeting the activation of immune checkpoints is recognized as an effective strategy for triggering anti-tumour immune responses in cancer cells. Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) were identified as potential crucial targets for cancer t...

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Main Authors:	Ronny Sham, Nur Fatihah, Abdul Hamid Hasani, Narimah, Idorus, Mohd Yusri, Abdul Karim, Muhammad Khalis, Syed Ahmad Fuad, Syed Baharom, Hasbullah, Harissa Husainy, Ibahim, Mohammad Johari
Format:	Article
Published:	Penerbit UTM Press 2023
Online Access:	http://psasir.upm.edu.my/id/eprint/110114/ https://mjfas.utm.my/index.php/mjfas/article/view/2700
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spelling	my.upm.eprints.1101142024-09-05T07:16:19Z http://psasir.upm.edu.my/id/eprint/110114/ Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot Ronny Sham, Nur Fatihah Abdul Hamid Hasani, Narimah Idorus, Mohd Yusri Abdul Karim, Muhammad Khalis Syed Ahmad Fuad, Syed Baharom Hasbullah, Harissa Husainy Ibahim, Mohammad Johari Targeting the activation of immune checkpoints is recognized as an effective strategy for triggering anti-tumour immune responses in cancer cells. Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) were identified as potential crucial targets for cancer treatment. Overexpression of CTLA-4 and PD-1 proteins in primary tumour and human cell lines is well documented. In contrary, lack of data was available using animal cell lines. The presence study aims to optimize the expression of CTLA-4 and PD-1 proteins in EMT6 mouse mammary cancer cells using Western blot, and provide basic understanding of their association with breast cancer cell progression. Proteins extracted from EMT6 parental cells were adjusted to 30ng for gel electrophoresis. Afterwards, the protein was transferred to a nitrocellulose membrane for blotting. The membrane was then subjected to chemiluminescent for band detection. Results obtained using beta-actin as a housekeeping gene show that both CTLA-4 (32 kDa) and PD-1 (34 kDa) proteins were expressed by using a 1:1000 dilution for each antibody from the lysate of EMT6 mouse mammary cancer cells. The relative expression of PD-1 (4.0 ± 0.26) is higher compared to CTLA-4 (1.2 ± 1.8). As a conclusion, both CTLA-4 and PD-1 proteins were indeed expressed in EMT6 mouse mammary cancer cells and this outcome provide the platform for extensive in vivo research on the link of both proteins with breast cancer using animal model. Penerbit UTM Press 2023 Article PeerReviewed Ronny Sham, Nur Fatihah and Abdul Hamid Hasani, Narimah and Idorus, Mohd Yusri and Abdul Karim, Muhammad Khalis and Syed Ahmad Fuad, Syed Baharom and Hasbullah, Harissa Husainy and Ibahim, Mohammad Johari (2023) Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot. Malaysian Journal of Fundamental and Applied Sciences, 19. pp. 194-201. ISSN 2289-5981; ESSN: 2289-599X https://mjfas.utm.my/index.php/mjfas/article/view/2700 10.11113/mjfas.v19n2.2700
institution	Universiti Putra Malaysia
building	UPM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Putra Malaysia
content_source	UPM Institutional Repository
url_provider	http://psasir.upm.edu.my/
description	Targeting the activation of immune checkpoints is recognized as an effective strategy for triggering anti-tumour immune responses in cancer cells. Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) were identified as potential crucial targets for cancer treatment. Overexpression of CTLA-4 and PD-1 proteins in primary tumour and human cell lines is well documented. In contrary, lack of data was available using animal cell lines. The presence study aims to optimize the expression of CTLA-4 and PD-1 proteins in EMT6 mouse mammary cancer cells using Western blot, and provide basic understanding of their association with breast cancer cell progression. Proteins extracted from EMT6 parental cells were adjusted to 30ng for gel electrophoresis. Afterwards, the protein was transferred to a nitrocellulose membrane for blotting. The membrane was then subjected to chemiluminescent for band detection. Results obtained using beta-actin as a housekeeping gene show that both CTLA-4 (32 kDa) and PD-1 (34 kDa) proteins were expressed by using a 1:1000 dilution for each antibody from the lysate of EMT6 mouse mammary cancer cells. The relative expression of PD-1 (4.0 ± 0.26) is higher compared to CTLA-4 (1.2 ± 1.8). As a conclusion, both CTLA-4 and PD-1 proteins were indeed expressed in EMT6 mouse mammary cancer cells and this outcome provide the platform for extensive in vivo research on the link of both proteins with breast cancer using animal model.
format	Article
author	Ronny Sham, Nur Fatihah Abdul Hamid Hasani, Narimah Idorus, Mohd Yusri Abdul Karim, Muhammad Khalis Syed Ahmad Fuad, Syed Baharom Hasbullah, Harissa Husainy Ibahim, Mohammad Johari
spellingShingle	Ronny Sham, Nur Fatihah Abdul Hamid Hasani, Narimah Idorus, Mohd Yusri Abdul Karim, Muhammad Khalis Syed Ahmad Fuad, Syed Baharom Hasbullah, Harissa Husainy Ibahim, Mohammad Johari Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
author_facet	Ronny Sham, Nur Fatihah Abdul Hamid Hasani, Narimah Idorus, Mohd Yusri Abdul Karim, Muhammad Khalis Syed Ahmad Fuad, Syed Baharom Hasbullah, Harissa Husainy Ibahim, Mohammad Johari
author_sort	Ronny Sham, Nur Fatihah
title	Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
title_short	Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
title_full	Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
title_fullStr	Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
title_full_unstemmed	Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot
title_sort	optimization of ctla-4 and pd-1 proteins in emt6 mouse mammary cancer cells by western blot
publisher	Penerbit UTM Press
publishDate	2023
url	http://psasir.upm.edu.my/id/eprint/110114/ https://mjfas.utm.my/index.php/mjfas/article/view/2700
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score	13.211869

Optimization of CTLA-4 and pd-1 proteins in EMT6 mouse mammary cancer cells by Western blot

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