Figure 1: Treatment of NIH/3T3 cells with various concentrations of LY294002 blocks PDGF-stimulated phosphorylation of Akt1 at serine 473 detected by PathScan Phospho-Akt1 (Ser473) Sandwich ELISA kit , #7160, but does not affect the level of total Akt1 protein detected by PathScan Total Akt1 Sandwich ELISA kit #7170. OD450 readings are shown in the top figure, while the corresponding Western blot, using Phospho-Akt (Ser473) Antibody #9271 or Akt Antibody #9272, is shown in the bottom figure.
Figure 4: Linear relationship between protein concentration of lysates from either control NIH/3T3 cells (dotted line) or PDGF-treated NIH/3T3 cells (solid line) and kit assay optical density readings.
Figure 3: Comparison of sensitivity between sandwich ELISA and Western blot in the detection of recombinant nonphospho-Akt1 protein, using Akt Antibody #9272. The top figure indicates that, at 0-1,000 pg of protein, the concentration of nonphospho-Akt1 is linearly proportional to the OD450 reading from the Sandwich ELISA kit.
Figure 2: Comparison of sensitivity between sandwich ELISA and Western blot in the detection of recombinant phospho-Akt1 protein, using Akt Antibody #9272. The top figure indicates that, at 0-500 pg of protein, the concentration of phospho-Akt1 is linearly proportional to the OD450 reading from the Sandwich ELISA kit.