Anti mouse igg1 alexa fluor 594

Cryosections of 10 μm were cut from frozen muscle using the Microm HM 550. Cryosections were rinsed once with PBS and fixed in 2% PFA in PBS for 5 min at RT. Sections were rinsed three times for 5 min with PBS, permeabilized with 0.5 % Triton X-100/0.1 M Glycine in PBS for 5 min at RT followed again by rinsing them three times with PBS. Sections were blocked in PBS supplemented with 5 % Horse serum and 1:40 Mouse on mouse blocking reagent (Vector labs) for 1 h at RT. Incubation with primary antibodies was carried out overnight at 4°C. The next day, sections were rinsed three times with PBS followed by incubation with secondary antibodies for 1 h at RT. Sections were rinsed again with PBS and nuclei were counterstained with 1:1,000 DAPI/PBS before mounting with Permaflour (Thermo Scientific). Slides were stored at 4°C until analysis. The following primary antibodies were used: 1:1,000 chicken anti-GFP (ab6556, AbCam), 1:1,000 rabbit anti-Laminin (L9393, Sigma), 1:200 rabbit anti-Ki67 (ab15580, AbCam), undiluted mouse anti-Pax7 (DSHB). The following secondary antibodies were used at 1:1,000 concentration: anti-chicken IgG Alexa-Fluor 488, anti-rabbit IgG Alexa-Fluor 488, anti-mouse IgG1 Alexa-Fluor 594 (Life Technologies).