Fig 1: Effect of buffer reagent on Western blotting linearity.(A) Western blot of rat liver samples (3–12 µg) using anti-PSMA6 and different buffers (TBST and PBST). (B) PSMA6 quantification, not normalized to total protein. (C) PSMA6 quantification, normalized to total protein. (D) Western blot of rat liver samples (3–12 µg) using anti-ß-actin and different buffers. (E) ß-actin quantification, not normalized to total protein. (F) ß-actin quantification, normalized to total protein. * p < 0.05, ** p < 0.01 by 1-way ANOVA.
Fig 2: Effect of antibody concentration on linearity of target proteins detected by Western blotting.(A) Western blot of rat liver samples (3–12 µg) using anti-PSMA6 at four different concentrations (1:5000, 1:10000, 1:20000, and 1:50000). (B) Quantification of anti-PSMA6 Western blots without including any normalization. (C) Quantification of anti-PSMA6 Western blots using total protein normalization. (D) Western blot of rat liver samples (3–12µg) using anti-ß-actin at four different concentrations (1:1000, 1:2500, 1:10000, and 1:25000). (E) Quantification of anti-ß-actin Western blots without including any normalization. (F) Quantification of anti-ß-actin Western blots using total protein normalization. * p < 0.05 by 1-way ANOVA.
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