Fig 1: Example of a Western blotting experiment that could be used to estimate differences in protein abundance. (a) Fluorescent image of the membrane showing proteins in each lysate that were labelled with Cy5. SeeBlue Plus2 Marker was used for sizing (not shown). This image was used to estimate the total protein abundance for each sample. Arrows indicate the approximate location of ENPP1 (open arrow) and Fam3a (closed arrow). (b) Western blot images of recombinant ENPP1, and Fam3a that were used to compare their abundance. These proteins were detected by chemiluminescence. (c) The standard curve of recombinant ENPP1 O.D. values that was used to interpolate levels of recombinant ENPP1 in each sample. (d) The standard curve of recombinant Fam3a O.D. values that was used to interpolate levels of recombinant Fam3a in each sample. (e) This table shows the results of interpolating the O.D. data against the standard curve for that protein as well as the comparisons made using the raw O.D. data.
Fig 2: Densitometry analyses and representative Western blots of lysates spiked with recombinant proteins. (a) Representative blot detected using chemiluminescence. (b) Representative blot detected using infrared fluorescence. In both images the top band at approximately 120 kDa is recombinant ENPP1 (open arrow) and the bottom band at approximately 25 kDa is recombinant Fam3a (closed arrow). MagicMark XP was used for sizing (not shown). (c) ENPP1 O.D. values in membranes detected with chemiluminescence. (d) ENPP1 O.D. values in membranes detected with infrared fluorescence. (e) Fam3a O.D. values in membranes detected with chemiluminescence. (f). Fam3a O.D. values in membranes detected with infrared fluorescence. 3 independently prepared membranes were used for each detection method.
Supplier Page from Abcam for Recombinant Human Fam3a protein