To my knowledge the company BIOMAKOR later on went to become part of SIGMA.ĭue to the prevalence of anti-actin antibodies in Western blot as a loading control and the prevalence of Western blot analysis in the overall body of literature, this beta-actin clone is likely the most cited antibody clone among all antibody clones in the literature. Lando and BIOMAKOR generously granted us the publication of 2 initial papers in 1994 and provided us with a generous supply of the AC-15 Ab for many years. The probe demonstrated excellent specificity and worked well in all techniques and fixation methods used.ĭr. Joel Vandekerckhove from the University of Ghent, Belgium, we characterized the Ab by light and electron microscopy, as well as 1D and 2D Western Blotting (on whole cell lysates and on isoform-specific peptides). Alison North in Salzburg, and together with our collaborator, the actin isoform expert Prof. I was given the chance to take some of the putative beta-actin specific probes (AC 15 and AC 74) back to Salzburg to test the Ab in our then lab in Salzburg. Zeev Lando of BIOMAKOR instructed me on their latest developments that included isoform-specific actin antibodies (at that time excellent probes for muscle actins and pan-actin Abs were available, but a beta-isoform-specific probe was lacking). While conducting experiments in 1993 at the Weizmann Institute of Science in Rehovot, Israel, I was invited to visit the facilities of BIOMAKOR, a company focusing on the development of monoclonal antibodies. Small lab has kindly provided the following account of history about the clone AC-15 to this article. This clone is available from most antibody suppliers.ĭr. The clone specifically targets the beta isoform, with no cross-reactivity against the other 5 isoforms of actin family. AC-15 belongs to the subclass IgG1, and was raised in mice against the N-terminal synthetic peptide of beta-actin, with the sequence of AcD-D-D-I-A-A-L-V-I-D-N-G-S-G-K coupled to keyhole limpet hemocyanin. Victor Small's lab at the Institute for Molecular Biology, Austrian Academy of Sciences in their 1994 publications. ĪC-15 clone was characterized and reported by Dr. Other commonly used clones include AC-74 (MilliporeSigma A5316 and A2228 ) and C4 (SCBT sc-47778 MilliporeSigma MAB1501R). Its specificity has been confirmed in knockout studies. The most cited anti-beta-actin antibody clone is AC-15, which was used in Western blot, for example, in order to normalize Western blot signals for quantitative measurements and intein-mediated protein trans-splicing.

Anti actin antibody western blot