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. 2021 Jan 15;11:1491. doi: 10.1038/s41598-020-80402-1

Figure 2.

Figure 2

Coomassie blue stained gel (a) and Western immunoblotting (b) of the EPO proteins. EPO standard, wild type and mutant proteins were separated according to their molecular mass in 12% SDS polyacrylamide gel at a constant 175 voltages for 1.5 h. (a) The purity of separated proteins was visualized by Coomassie blue staining. The gel image was taken using HP LaserJet scanner. (b) The identity of EPO antigen was detected by Western immunoblotting. The chemiluminescence signals of protein samples were detected and the image was taken using ImageQuant LAS 4000 control software version 1.2 provided with ImageQuant LAS 4000 machine at the exposure time of 1 s. The signals were then overlaid with the marker using ImageQuant TL software version 7.0 (https://www.cytivalifesciences.com/en/us). The native gel and blot are presented in Supplementary Figs. S1 and S2, respectively. BRP, EPO standard; Marker, Precision Plus Protein Kaleidoscope Prestained Protein Standards; WT, EPO-WT; 1.2, EPO-1.2 (L70V, V74L, L102I); 3.1, EPO-3.1 (T106A); 3.2, EPO-3.2 (T106G); 3.3, EPO-3.3 (T106H); 4.1, EPO-4.1 (L109A).