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. 2022 Jun 16;13:918100. doi: 10.3389/fphys.2022.918100

FIGURE 9.

FIGURE 9

Validation of anti-nucleotidase antibodies used for enzyme identification. ProteinSimple Wes was used to specifically detect the indicated nucleotidases in (A) mouse brain homogenates (positive controls) and in (B) mouse tissue homogenates in which the indicated nucleotidase has not been detected (negative controls). Panel (Ba–Bi) show representative blot images (left) and immunoelectropherograms (right) of negative control tissue homogenates. Each antibody was tested in duplicate. The antibodies, dilutions, corresponding amounts of homogenate per well, and expected molecular weight (from vendor) used were: rabbit anti-ENTPD1 (1:200, 15 μg, and 80 kDa), sheep anti-ENTPD2 (1:500, 7.5 µg, and 80 kDa); rabbit anti-ENTPD3 (1:500, 3.0 µg, and 80 kDa); rabbit anti-ENTPD8 (1:200, 15 μg, and 60 kDa); rabbit anti-ENPP1 (1:500, 15 μg, and 100 kDa); rabbit anti-ENPP3 (1:500, 15 μg, and 100 kDa); rabbit anti-NT5E (1:500, 15 μg, 7and 0 kDa); rabbit anti-TNAP (1:200, 30 μg, and 60 kDa; rabbit anti-NT5C1A (1:100, 30 μg, and 40 kDa)).