A Schematic illustration of the PentaTag Control Protein DYKDDDDK-HA-MBP-Myc-ALFA (NanoTag #P0102). B SDS-PAGE analysis of 2 µg of PentaTag Control Protein per lane. The gel was stained with Coomassie Blue. C Western blot analysis of PentaTag Control Protein using 10 or 100 ng of recombinant protein per lane. The blots were probed as follows: DYKDDDDK: 1. anti-FLAG M2 (Sigma #F1804) 1:1000; 2. FluoTag®-X2 anti-Mouse IgG1 Atto643 (NanoTag #N2002) 1:5000; HA: 1. anti-HA Clone F-7 (Santa Cruz #sc-7392) 1:200, 2. FluoTag®-X2 anti-Mouse IgG2a/b Atto643 (NanoTag #N2702) 1:5000; MBP: 1. anti-MBP (Synaptic Systems #416 003) 1:5000, 2. FluoTag-X4 anti-Rabbit IgG Atto643 (NanoTag #N2404) 1:5000; Myc: 1. anti-Myc Clone 9E10 (Synaptic Systems #343 011) 1:1000, 2. FluoTag®-X2 anti-Mouse IgG1 Atto643 (NanoTag #N2002) 1:5000, ALFA: FluoTag®-X2 anti-ALFA Atto643 (NanoTag #N1502) 1:5000. All blots were analyzed using fluorescent detection with an excitation wavelength of 643 nm and emission at 665 nm.

Pulldown of PentaTag Control Protein DYKDDDDK-HA-MBP-Myc-ALFA (Cat. No. P0102) from HeLa lysate using ALFA Selector^CE (Cat. No. N1512). A HeLa lysate containing a defined amount of PentaTag Control Protein was used as input material. Proteins bound to ALFA Selector^CE were eluted using ALFA elution peptide (Cat. No. N1520). Indicated protein fractions were analyzed by SDS-PAGE followed by Coomassie Blue staining.