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A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.

Zhang Z, Chen J, Guo F, He L, Wu Y, Zeng C, Xiao X, He D - PLoS ONE (2008)

Bottom Line: A significant number of differential proteins can be discovered or re-categorized by this technique.Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively.Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

View Article: PubMed Central - PubMed

Affiliation: Universities' Confederated Institute of Proteomics, Key laboratory for Cell Proliferation and Regulation Biology Ministry of Education, Beijing Normal University, Beijing, People's Republic of China.

ABSTRACT
As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a (35)S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

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Related in: MedlinePlus

Comparison among BrdU labeling, Flow Cytometry and SiLAD to check cell cycle stage.Black line a means only to determine the cell cycle phase roughly. Curve line b means detecting the specific time point exactly. BrdU L is stands for the BrdU labeling and FCM is Flow Cytometry.
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pone-0002991-g005: Comparison among BrdU labeling, Flow Cytometry and SiLAD to check cell cycle stage.Black line a means only to determine the cell cycle phase roughly. Curve line b means detecting the specific time point exactly. BrdU L is stands for the BrdU labeling and FCM is Flow Cytometry.

Mentions: In this study, the SiLAD bar code was compared with two alternative methods, BrdU labeling and Flow Cytometry, which are also used to provide the temporal coordinate when a continuous bio-process is investigated. With BrdU labeling, an experienced researcher can easily discriminate four or five stages of S phase in cell cycle based on the characteristic spatial patterns of nuclear staining [15]. Flow cytometry provides the information to distinguish the whole G1 or G2/M phase and different stages within S phase. SiLAD bar code, in stead, can be used to determine all the temporal positions throughout the whole biological procession, yet with much higher resolution. This comparison was summed in the Figure 5. However, this SiLAD coordinate system has to be defined individually for each desired bio-process prior to use.


A high-temporal resolution technology for dynamic proteomic analysis based on 35S labeling.

Zhang Z, Chen J, Guo F, He L, Wu Y, Zeng C, Xiao X, He D - PLoS ONE (2008)

Comparison among BrdU labeling, Flow Cytometry and SiLAD to check cell cycle stage.Black line a means only to determine the cell cycle phase roughly. Curve line b means detecting the specific time point exactly. BrdU L is stands for the BrdU labeling and FCM is Flow Cytometry.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2500177&req=5

pone-0002991-g005: Comparison among BrdU labeling, Flow Cytometry and SiLAD to check cell cycle stage.Black line a means only to determine the cell cycle phase roughly. Curve line b means detecting the specific time point exactly. BrdU L is stands for the BrdU labeling and FCM is Flow Cytometry.
Mentions: In this study, the SiLAD bar code was compared with two alternative methods, BrdU labeling and Flow Cytometry, which are also used to provide the temporal coordinate when a continuous bio-process is investigated. With BrdU labeling, an experienced researcher can easily discriminate four or five stages of S phase in cell cycle based on the characteristic spatial patterns of nuclear staining [15]. Flow cytometry provides the information to distinguish the whole G1 or G2/M phase and different stages within S phase. SiLAD bar code, in stead, can be used to determine all the temporal positions throughout the whole biological procession, yet with much higher resolution. This comparison was summed in the Figure 5. However, this SiLAD coordinate system has to be defined individually for each desired bio-process prior to use.

Bottom Line: A significant number of differential proteins can be discovered or re-categorized by this technique.Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively.Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

View Article: PubMed Central - PubMed

Affiliation: Universities' Confederated Institute of Proteomics, Key laboratory for Cell Proliferation and Regulation Biology Ministry of Education, Beijing Normal University, Beijing, People's Republic of China.

ABSTRACT
As more and more research efforts have been attracted to dynamic or differential proteomics, a method with high temporal resolution and high throughput is required. In present study, a (35)S in vivo Labeling Analysis for Dynamic Proteomics (SiLAD) was designed and tested by analyzing the dynamic proteome changes in the highly synchronized A549 cells, as well as in the rat liver 2/3 partial hepatectomy surgery. The results validated that SiLAD technique, in combination with 2-Dimensional Electrophoresis, provided a highly sensitivity method to illustrate the non-disturbed endogenous proteins dynamic changes with a good temporal resolution and high signal/noise ratio. A significant number of differential proteins can be discovered or re-categorized by this technique. Another unique feature of SiLAD is its capability of quantifying the rate of protein expression, which reflects the cellular physiological turn points more effectively. Finally, the prescribed SiLAD proteome snapshot pattern could be potentially used as an exclusive symbol for characterizing each stage in well regulated biological processes.

Show MeSH
Related in: MedlinePlus