Anti alexafluor 488 antibody

C57Bl/6 mice were injected bilaterally with CTB 488 into the colon as described above. NG were dissected 1 week post injection as described above, dipped in Fast Green (1%, Sigma-Aldrich) to assist with visualization and flash frozen in OCT. 15 um sections of NG were sliced on a cryostat for RNAScope/IHC. Samples were processed and stained with Scn5a, positive or negative control probes according to the manufacturer’s instructions. After in situ hybridization sections were washed three times in wash buffer (1X, ACDBio) and then fixed in 1% PFA in TBS for 10 minutes at 4 C to stabilize the ISH labeling. Samples were next washed three times in TBS-T and incubated in 10% Goat Serum in TBS with 1% BSA for 30 minutes. Samples were stained with anti Alexafluor-488 antibody (1:1000, Thermo-Fisher) for 1 hour in TBS-1% BSA. After primary antibody staining, sections were washed three times for 5 minutes each in TBST and stained with Goat anti rabbit AF488 (1:1000, Thermo-Fisher) in TBS-1% BSA for 30 minutes. Samples were again washed three times for 5 minutes each in TBST and finally mounted in Prolong gold antifade with DAPI (Thermo-Fisher) for imaging. Slides were imaged within 24 hours of mounting on an inverted LSM 880 NLO laser scanning confocal and multiphoton microscope (Zeiss) and images were processed using Image J.