𝔖 Bobbio Scriptorium
✦   LIBER   ✦

A novel flow cytometric method for the quantification of p53 gene expression

✍ Scribed by Giuditta Filippini; Sayuri Griffin; Mario Uhr; Hans Eppenberger; Josè Bonilla; Franco Cavalli; Gianni Soldati

Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
87 KB
Volume
31
Category
Article
ISSN
0196-4763

No coin nor oath required. For personal study only.

✦ Synopsis

We describe a rapid and simple flow cytometry technique for the detection and quantification of p53 in several human cell lines, including an adenocarcinoma cell line (SW 626) having a mutant (m) p53, and a pre-B leukemia cell line (NALM-6) having wild-type (wt) p53. By introducing a second antibody coupled to RPE-fluorescence, the discrimination between control and specific peaks was improved over that achieved with methods used previously. To quantify the content of p53 molecules in the cells, we used a series of beads with the capability to bind mouse monoclonal IgG antibodies. p53 cell content, expressed as antibody binding capacity (ABC), was directly quantified from logarithmic scattergrams; the results were reproducible in all cell lines tested. Flow cytometric results were compared with those of a standard immunocytochemistry method routinely used for the detection of p53 in cells, and found to be in correlation. Furthermore, cytometric data also reflect ELISA determinations. In summary, we showed for the first time that (i) p53 can be clearly detected by flow cytometry in various cell lines, (ii) p53 can be quantitated in terms of number of molecules per cell, and (iii) it can be easily monitored as a function of time after stimulation.

📜 SIMILAR VOLUMES

A novel flow cytometric method for quant
A novel flow cytometric method for quantifying phagocytosis of apoptotic cells
✍ Krista L. Hess; George F. Babcock; David S. Askew; Joan M. Cook-Mills 📂 Article 📅 1997 🏛 John Wiley and Sons 🌐 English ⚖ 235 KB 👁 2 views

Many eukaryotic cell types are capable of specific recognition and phagocytosis of apoptotic cells, and there is increasing interest in the mechanisms involved in this process. To facilitate analysis of these mechanisms, we designed a novel fluorescence-based method to quantify phagocytosis in vitro

Evaluation of a flow cytometric method f
Evaluation of a flow cytometric method for simultaneous leukocyte phenotyping and quantification by fluorescent microspheres
✍ P. Schlenke; C. Frohn; H. Klüter; M. Saballus; H. J. Hammers; S. R. Zajac; H. Ki 📂 Article 📅 1998 🏛 John Wiley and Sons 🌐 English ⚖ 120 KB 👁 1 views

We describe a flow cytometric method using a newly designed product, fluorochrome-containing microspheres (Flow Count fluorospheres), which facilitates the precise quantification of cells in whole blood samples or heterogeneous cell suspensions on a single-cell level. These microparticles are easily

Attachment staining: A novel method for
Attachment staining: A novel method for flow cytometric quantitation of protein expression in limited numbers of adherent cells
✍ Shaun R. Hawley; Stephen R. Pennington 📂 Article 📅 1998 🏛 John Wiley and Sons 🌐 English ⚖ 164 KB 👁 1 views

Measurement of protein expression in adherent cells in culture by flow cytometry, although potentially a very powerful method, has so far found limited application. Where the method has been applied successfully, cells have been resuspended for analysis either prior to experimental treatment or befo

Comparison of flow cytometry and laser s
Comparison of flow cytometry and laser scanning cytometry for the intracellular evaluation of adenoviral infectivity and p53 protein expression in gene therapy
✍ Mary L. Musco; Shijun Cui; David Small; Margarita Nodelman; Barry Sugarman; Mich 📂 Article 📅 1998 🏛 John Wiley and Sons 🌐 English ⚖ 127 KB 👁 1 views

The determination of recombinant adenoviral (rAd) infectivity and p53 protein expression is important for the evaluation of rAd vectors containing the p53 gene (rAd-CMV-p53) for gene therapy. We have previously reported that rAd5-CMV-p53 vectors can be assessed for infectivity and concomitant p53 pr