As presented in Fig. 2A, in control groups, no specific blot was detected while in the overexpression group, a clear Myc6-Bub3 blot could be observed (40 (Bub3) kDa + approximate 10 (Myc) kDa = approximate 50 kDa), indicating the expression of Myc6-Bub3 mRNA.
Equal loading was confirmed with specific blotting for ERK2, which can be done without stripping.
A faint band (Mr ≈ 120K, labelled c) was observed on some blots due to specific blotting conditions.
We observed that many commercially available antibodies, including H-300, P20, and 8G7, exhibited marginal specific blotting and/or multiple background bands (not shown).
When taking into account the lack of specificity of a number of the immunoblots, it is clear that the combination of a non-specific ELISA with a non-specific blot will lead to a high fraction of presumably false-positive test results.
PrP-specific Western blots revealed three PrP specific signals from 25 to 35 kDa in the cells expressing PrP-WT, which represented di- mono- and un-glycosylated PrP.
Two of the independent methods employed are quantitative and Aβ specific (Western blots and immunostaining).
HIV testing was done by enzyme linked immunosorbent assay (ELISA; Wellcome Laboratories, Dartford, Kent, UK) and by type-specific Western blot (New LAV Blot I and New LAV Blot II, Diagnostics Pasteur, Marnes-la-Coquette, France).
Results from the kinome analysis were validated by performing phospho-specific Western blots for major kinases.
Following treatment, whole-cell lysates were subjected to specific Western blot analyses.
