Mouse anti mmp1 c terminus

Larvae were dissected in 1 x PBS followed by a 20-min fix in 4% paraformaldehyde in PBS. After 3 washes in 0.1% PBST (1 x PBS + 0.1%Triton-X), larvae were washed in 0.3% PBST and then blocked in 0.1% PBST with 5% normal goat serum (NGS) for 30 min. Primary staining was done overnight at 4°C, whereas secondary staining was done for 4 h at room temperature. The following primary antibodies were obtained from the Developmental Studies Hybridoma Bank: mouse anti-Nubbin (1:25), mouse anti-Wg (1:100), mouse anti-Mmp1 C-terminus (1:100), mouse anti-Mmp1 catalytic domain (1:100), mouse anti-LacZ (1:100), and rat anti-DE-cadherin (1:100). Rabbit anti-DCP1 (1:1000), and mouse anti-PH3 (1:400) were obtained from Cell Signaling Technologies, and chick anti-GFP (1:2000) was obtained from Abcam. The secondary anti-Chick 488 antibody was also obtained from Abcam. Other secondary antibodies, anti-rabbit 647, anti-rat 647, and anti-mouse 555 were obtained from Invitrogen. All secondary fluorophore-conjugated antibodies were used at 1:500. Images were obtained on a Zeiss AxioImager.M2 with ApoTome. For each experiment at least 15 discs were analyzed prior to choosing a representative image. Images were processed using Affinity Photo and Affinity Designer.