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A rapid microwell fluorescence immunoassay for cellular protein detection.

Lavigne C, Guignée de A, Thierry AR - Biol Proced Online (2008)

Bottom Line: In order to illustrate this method, we report on the detection of two different proteins, the cell cycle proteins cyclin D1 and p21(CIP1/WAF1), in untreated and 2-cyclopenten-1-one treated breast cancer cells.When the FICP method was compared with Western blot procedure, FICP was found to be superior for many characteristics.By using this method, we were able to quantify biological effects of a specific compound on protein levels in non-lysed cells and perform statistical analysis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Beausejour Medical Research Institute, Moncton, New Brunswick, Canada. lavigne@phac-aspc.gc.ca

ABSTRACT
In this paper, we describe a simple, rapid, specific, sensitive, and reliable method, the FICP method (Fluorescence Immunoassay for Cellular Protein detection) which is readily applicable to the detection of proteins directly on cells cultured in 96-well plates. In order to illustrate this method, we report on the detection of two different proteins, the cell cycle proteins cyclin D1 and p21(CIP1/WAF1), in untreated and 2-cyclopenten-1-one treated breast cancer cells. When the FICP method was compared with Western blot procedure, FICP was found to be superior for many characteristics. By using this method, we were able to quantify biological effects of a specific compound on protein levels in non-lysed cells and perform statistical analysis. Therefore, we believe this screening assay could be very useful for detecting poorly expressed proteins and for drug development.

No MeSH data available.


Related in: MedlinePlus

Effect of cyclopentenone on MCF-7  cell  proliferation.				Cells were plated in 96- well plates at a density of 10,000 cells per well. Viable cells were measured using the MTT Cell Proliferation Assay 4 hours after treatment with increasing doses of cyclopentenone. Data shown represent mean of 12 replicates ± SD from two separate experiments, each done in 6 replicates.
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Figure 03: Effect of cyclopentenone on MCF-7 cell proliferation. Cells were plated in 96- well plates at a density of 10,000 cells per well. Viable cells were measured using the MTT Cell Proliferation Assay 4 hours after treatment with increasing doses of cyclopentenone. Data shown represent mean of 12 replicates ± SD from two separate experiments, each done in 6 replicates.

Mentions: Since cydopentenone causes an arrest in cell cyde and inhibits tumor cells proliferation (17-19) it was important to assess whether the diminution in the production of cydin D1 observed in the fluorescence immunoassay and the Western Blot method was correlated with an inhibition in the proliferation of the cells. As shown inFig. 3, the number of cells decreased in a dose-response manner with increasing concentration of cyclopentenone in the culture medium.


A rapid microwell fluorescence immunoassay for cellular protein detection.

Lavigne C, Guignée de A, Thierry AR - Biol Proced Online (2008)

Effect of cyclopentenone on MCF-7  cell  proliferation.				Cells were plated in 96- well plates at a density of 10,000 cells per well. Viable cells were measured using the MTT Cell Proliferation Assay 4 hours after treatment with increasing doses of cyclopentenone. Data shown represent mean of 12 replicates ± SD from two separate experiments, each done in 6 replicates.
© Copyright Policy - open acces
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2683549&req=5

Figure 03: Effect of cyclopentenone on MCF-7 cell proliferation. Cells were plated in 96- well plates at a density of 10,000 cells per well. Viable cells were measured using the MTT Cell Proliferation Assay 4 hours after treatment with increasing doses of cyclopentenone. Data shown represent mean of 12 replicates ± SD from two separate experiments, each done in 6 replicates.
Mentions: Since cydopentenone causes an arrest in cell cyde and inhibits tumor cells proliferation (17-19) it was important to assess whether the diminution in the production of cydin D1 observed in the fluorescence immunoassay and the Western Blot method was correlated with an inhibition in the proliferation of the cells. As shown inFig. 3, the number of cells decreased in a dose-response manner with increasing concentration of cyclopentenone in the culture medium.

Bottom Line: In order to illustrate this method, we report on the detection of two different proteins, the cell cycle proteins cyclin D1 and p21(CIP1/WAF1), in untreated and 2-cyclopenten-1-one treated breast cancer cells.When the FICP method was compared with Western blot procedure, FICP was found to be superior for many characteristics.By using this method, we were able to quantify biological effects of a specific compound on protein levels in non-lysed cells and perform statistical analysis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Beausejour Medical Research Institute, Moncton, New Brunswick, Canada. lavigne@phac-aspc.gc.ca

ABSTRACT
In this paper, we describe a simple, rapid, specific, sensitive, and reliable method, the FICP method (Fluorescence Immunoassay for Cellular Protein detection) which is readily applicable to the detection of proteins directly on cells cultured in 96-well plates. In order to illustrate this method, we report on the detection of two different proteins, the cell cycle proteins cyclin D1 and p21(CIP1/WAF1), in untreated and 2-cyclopenten-1-one treated breast cancer cells. When the FICP method was compared with Western blot procedure, FICP was found to be superior for many characteristics. By using this method, we were able to quantify biological effects of a specific compound on protein levels in non-lysed cells and perform statistical analysis. Therefore, we believe this screening assay could be very useful for detecting poorly expressed proteins and for drug development.

No MeSH data available.


Related in: MedlinePlus