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This video demonstrates the methodology for performing Reverse Phase Protein Arrays (RPPA) to study protein expression patterns from cell lysate. In RPPA, the lysate containing a mixture of proteins is printed on nitrocellulose slides and proteins of interest are analyzed using fluorescently labeled antibodies.
1. RPPA Printing
2. RPPA Protein Detection
Figure 1. Flow diagram of RPPA scanning process.
Name | Company | Catalog Number | Comments |
Triton X-100 | Triton-X | T8787 | |
Li-Cor Odyssey Blocking Buffer | Li-Cor | 927-40000 | |
MicroGrid II robotic spotter | Biorobotics | ||
FastFrame' four bay slide holder | Whatman | 10486001 | |
FAST Slide - 2-Pad | Whatman | 10485317 | |
IRDye 680LT Goat anti-Mouse IgG | Licor | 926-68020 | |
IRDye 800CW Goat anti-Rabbit IgG | Licor | 926-32211 |
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Source: O'Mahony, F. C. et al. The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers. J. Vis. Exp. (2013)
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