Figure 3.

Western blots with NeoMarkers TGase 2 antibodies of homogenate (H) and mitochondrial (M) fractions prepared from frozen liver obtained from KO and WT mice. H* and M* represent GTP-agarose pull downs from H and M fractions, respectively. The lane labeled TGase 2* contained TGase 2 that had been subjected to the GTP pull-down procedure. (A) Detection by the ECL procedure. Each lane, except that marked TGase 2* (which was loaded with 4 μg), contained 25 μg of protein. In this experiment, frozen liver was ground in dry ice and the powder was suspended in buffer and homogenized at 0 °C. The mitochondria were purified from this suspension. (B) Detection by the alkaline phosphatase procedure. Each lane except that marked TGase 2* (which was loaded with 8 μg), contained 80 μg of protein. In this experiment, the frozen liver was thawed and then homogenized in buffer at 0 °C and the mitochondria were prepared directly from this suspension. In this figure and in Figure 2, no immunopositive bands were detected in the low Mr range.