TY - JOUR
T1 - Scintillation proximity assay for total p53 protein as an alternative to ELISA
AU - Oberlander, Sarit
AU - Xie, Tian
AU - Chandrachud, Uma
AU - Gal, Susannah
N1 - Funding Information:
The work was supported by a National Institutes of Health grant ( R15 CA101783-01A1 ) and by the US Department of Energy (Grant DE-FG02-06ER64281 ) as a subcontract from SUNY-Utica both to SG. The work on developing the SPA for DNA binding was originally funded by Amersham Pharmacia Biotech, the original commercial source of the SPA beads used in this work. Since that time, the authors have had no financial support from the company or its new entity, GE Healthcare.
PY - 2010/8
Y1 - 2010/8
N2 - Measurement of the level of a specific protein can be an important parameter to discern as that can change and reflect disease status. A number of methods have been developed to quantitate the level of a protein, some amenable to high throughput screening. A method is described to measure the total level of the tumor suppressor p53 using scintillation proximity assay (SPA) beads and radiolabeled streptavidin. Three different cell extracts were used, with one used to develop the standard curve for the amount of p53. This method allows the specific detection of p53 in the range of 50 to 300. pg in 10 μl of an extract. While this detection is less than what can be detected by commercially available enzyme linked immunosorbent assay (ELISA) kits, the SPA compares favorably on time required and cost. This new assay also has the potential to be coupled with measurements for p53 DNA binding, a unique aspect of this approach.
AB - Measurement of the level of a specific protein can be an important parameter to discern as that can change and reflect disease status. A number of methods have been developed to quantitate the level of a protein, some amenable to high throughput screening. A method is described to measure the total level of the tumor suppressor p53 using scintillation proximity assay (SPA) beads and radiolabeled streptavidin. Three different cell extracts were used, with one used to develop the standard curve for the amount of p53. This method allows the specific detection of p53 in the range of 50 to 300. pg in 10 μl of an extract. While this detection is less than what can be detected by commercially available enzyme linked immunosorbent assay (ELISA) kits, the SPA compares favorably on time required and cost. This new assay also has the potential to be coupled with measurements for p53 DNA binding, a unique aspect of this approach.
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U2 - 10.1016/j.jim.2010.06.018
DO - 10.1016/j.jim.2010.06.018
M3 - Article
C2 - 20600074
AN - SCOPUS:77955663664
SN - 0022-1759
VL - 360
SP - 173
EP - 177
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -