Rabbit anti ldha

For whole-mount immunohistochemistry, embryos were collected at appropriate developmental stages and fixed in 4% PFA-PB for 20 mins at RT. Post fixation, embryos were dissected from the filter paper and washed in TBS containing 0.1% Triton and 1% DMSO (TBTD). Embryos were blocked at RT for 2 hours in TBTD supplemented with 10% donkey serum and incubated in primary antibody diluted in blocking solution, overnight at 4°C. The following primary antibodies were used: rabbit anti-PFKP (Abcam, 1:200), rabbit anti-LDHA (Abcam, 1:200), mouse anti-GAPDH (Millipore, 1:500), mouse anti-TFAP2B (Santa Cruz Biotechnology, 1:500), rabbit anti-ActYAP1 (Abcam, 1:500), rabbit anti-TFAP2B (Abcam, 1:250), rabbit anti-Sox9 (EMD Millipore, 1:500), mouse anti-E-Cadherin (BD Biosciences, 1:200). Following the primary antibody incubation, embryos were washed, blocked for 30 mins at RT, and stained with appropriate secondary antibodies for 2h at RT. Secondary antibodies used included donkey anti-mouse/rabbit IgG conjugated with Alexa Fluor 488/568/647 or goat anti-mouse Alexa 480/568 (Molecular Probes, 1:3000). Following the secondary antibody step, the embryos were washed, stained with Dapi and post-fixed with 4% PFA for 1h, prior to imaging. Whole-mount images were collected using an upright Zeiss Axio Imager fluorescent microscope and processed as described below.