Rabbit anti gs c 18 antibody

Samples from important steps during purification were collected and analysed by SDS-PAGE and western blot. TGX™ Precast Gel (BioRad) was used to separate proteins within samples at 200 V for 30 min. Then gels were either stained by Instant Blue (Sigma Aldrich) or immediately transferred to PVDF membrane (BioRad) at 100 V for 1 h. The proteins on the PVDF membrane were probed with two primary antibodies simultaneously, rabbit anti-Gs C-18 antibody (cat. no. sc-383, Santa Cruz) against Gαs subunit and mouse poly-His antibody (cat. no. 34660, QIAGEN) against His tags. The membrane was washed and incubated with secondary anti-mouse and anti-rabbit antibodies (LI-COR). The membranes were imaged on a Typhoon 5 imaging system (Amersham).

Samples from important steps during purification were collected and analysed by SDS-PAGE and western blot. TGX™ Precast Gel (BioRad) was used to separate proteins within samples at 200 V for 30 min. Subsequently, gels were either stained by Instant Blue (Sigma Aldrich) or immediately transferred to PVDF membrane (BioRad) at 100 V for 1 h. The proteins on the PVDF membrane were probed with two primary antibodies simultaneously, rabbit anti-Gs C-18 antibody (cat. no. sc-383, Santa Cruz) against Gαs subunit and mouse poly-His antibody (cat. no. 34660, QIAGEN) against His tags. The membrane was washed and incubated with secondary IRDye anti-mouse and IRDye anti-rabbit antibodies (LI-COR). The membranes were imaged at wavelengths of 680 and 800 nm to visualize His-tagged proteins (GLP-1R, Gβ1 and Nb35) and Gαs, respectively, on a Typhoon 5 imaging system (Amersham).