Alexa fluor 488 conjugated goat anti rabbit igg 111 545 144

Tctn3 antibody was generated by Covance. Rabbits were immunized with His-tagged mouse His-Tctn3(297-E) fragment purified from bacteria. This and other antibodies (1:1000 dilution, unless otherwise indicated) include: Gli2 N-terminal, Gli3 N-terminal (Pan et al., 2006 (link); Wang et al., 2000 (link)), ARL13B (Caspary et al., 2007 (link)), Smo (Rohatgi et al., 2007 (link)), IFT88 and IFT140 (Pazour et al., 2002 (link)), Cep164, Odf2, Ninein (Wu et al., 2014 ), AC3, Myc (sc-1701, sc-789, Santa Cruz Biotechnology), acetylated tubulin, FLAG (T6793, F9291, Sigma). Secondary antibodies Alexa Fluor 488-conjugated goat anti-rabbit IgG (111—545-144) and Cy3-conjugated goat anti-mouse IgG (115-165-146) were purchased from Jackson Immunoresearch, Inc.

Arl13b, AKAP9, Gli2, Gli3, and pGli antibodies were generated by Covance or Biosynthesis. Rabbits were immunized with purified bacterially expressed His-tagged mouse Arl13b, His-tagged mouse AKAP9 fragment (2768–3116 aa), His-tagged mouse Gli2C-terminal fragment (605–1460 aa), GST-tagged Gli3 C-terminal fragment (1051–1179 aa), or the synthetic Gli2P2 phospho-peptide (NH₂-CAYTVSRRS-(pS)-GISP-OH), in which residue C is not from Gli2 and was used for conjugation of the peptide to Keyhole Limpet Hemocyanin (KLH). It is worth noting that the sequence for Gli3 in the same region is AYLSSRRSSGISP with only the 3rd and 4th residues different from the Gli2. The phosphorylation site is conserved between Gli2 and Gli3. The antibodies were used in a 1:1000 dilution. Other antibodies include: Gli2 N-terminal, Gli3 N-terminal, Ta3, Cep120 (all 1:1000) (Pan et al., 2006 (link); Wang et al., 2000 (link); Wu et al., 2014 (link)), acetylated tubulin (1:4000), FLAG (T6793, F1804, Sigma), PKARIIβ (610625, BD Biosciences), and Myc (sc-788, Santa Cruz Biotechnology). Secondary antibodies Alexa Fluor 488-conjugated goat anti-rabbit IgG (111−545-144) and Cy3-conjugated goat anti-mouse IgG (115−165-146) were purchased from Jackson Immunoresearch, Inc.