Co-IP of MCM complex via pulldown of MCM6 using 5 µg of monoclonal anti-MCM6 IgG1 (Proteintech: 67989-1-Ig) and anti-mouse IgG1 VHH agarose (mIG1a). 5 µg of each IgG was spiked into HEK293T cell lysate derived from 1x10^7 cells. All subunits of the 600 kDa hetero-hexameric complex are successfully precipitated, as shown in a Coomassie blue stained SDS-gel and by Western blot analysis using subunit specific antibodies. Apparent molecular weights are provided. For input (IN) and flowthrough (FT) fractions, 1% was loaded, respectively. For bound (B) fraction and bound fraction of isotype control antibody (BISO Proteintech: 66360-1-Ig), 25% was loaded. Product codes for Proteintech-antibodies are provided in parentheses. Compared to Protein A agarose, mIG1a shows a superior precipitation of the MCM complex.

IP of PCNA by anti-mouse IgG1 VHH agarose (mIG1a) using a monoclonal PCNA antibody (Proteintech: 60097-1-Ig). As control, a IgG1 isotype control antibody (Proteintech: 66360-1-Ig) was used (BISO). 5 µg of each IgG was spiked into HEK293T cell lysate derived from 0.5x10^7 cells. 1% of input (IN) and flow through (FT) and 25% of bound (B) fraction was loaded onto an SDS-PAGE gel. For Western blot analysis PCNA was detected using a polyclonal rabbit IgG (Proteintech: 10205-2-AP) (1:5000) and an HRP-labeled goat anti-rabbit secondary (Proteintech: RGAR001). Compared to Protein A agarose, mIG1a shows a superior precipitation of PCNA.