Journal: BMC Genomics

Article Title: CRISPR/Cas9-mediated gene knockin in the hydroid Hydractinia symbiolongicarpus

doi: 10.1186/s12864-018-5032-z

Figure Lengend Snippet: Use of the StrepII-FLAG tag in immunoblots and pull-downs. a Western blot for FLAG on total protein lysates from Eef1a FLAG-P2A-tdTom transgenic and nontransgenic colonies. Sizes in italics are the top and bottom bands of the molecular size marker. b Western blot of StrepII tag on total protein lysates from the same colonies. The blot was treated to induce chemiluminescence and exposed to film for 5 min (left), followed by a second exposure for 60 min (right). The 15 kDa band suggests this antibody cross-reacts with some variants of an endogenous Hydractinia protein. Arrowhead indicates non-specific staining of protein ladder. c Coomassie stained SDS-PAGE gel of elutions from Strep-Tactin coated beads following incubation in total protein lysates from the indicated colonies. d Western blot against the FLAG peptide on the indicated amount of total protein lysate from colony 355–8, before and after incubation with the anti-streptavidin beads. The elution from the beads was run alongside for comparison

Article Snippet: The gel was then fixed for 10 min (10% acetic acid, 25% IPA) at RT, stained ON at RT in a 0.3% solution of G250 Brilliant Blue coomassie (BioRad 161–0406), destained with 10% acetic acid for approx.

Techniques: FLAG-tag, Western Blot, Transgenic Assay, Marker, Staining, SDS Page, Incubation, Comparison