MTHFR, Bethyl Laboratories, A303-624A

We generated a top 1000 list of the most studied and most published proteins and analyzed literature validation data for commercially available research antibodies. MTHFR encoding methylenetetrahydrofolate reductase is the eighth most studied gene on the list. In Li et al., Cell Cycle (2017), the authors investigate the role of MTHFR in DNA replication. Bethyl A303-624A is used for WB (1:1000) of lysates of HeLa cell following siRNA treatment (MTHFR and control). In Figure 3A, the antibody signal is substantially reduced with two independent MTHFR siRNAs compared with the control. In Figure 5A, the signal is restored by expression of siRNA-resistant Flag-tagged MTHFR. β-actin loading controls are shown, and anti-Flag detection provides additional confirmation of the expressed proteins. These data support specific detection of MTHFR in WB under the tested conditions. The blots are cropped and lack molecular weight markers, limiting assessment of band size and additional nonspecific bands. We have rated A303-624A based on the available information and according to the pAbmAbs criteria. We highly encourage other scientists to share their observations.