Journal: Histochemistry and cell biology

Article Title: Differential expression of PKCα and -β in the zebrafish retina.

doi: 10.1007/s00418-018-1764-8

Figure Lengend Snippet: Fig. 3 Western blot analysis of recombinant zebrafish cPKC proteins using PKCα and -β antibodies. Three different anti- bodies were used to compare the specificity of each antibody with recombinant zebrafish PKCs. The PKCα NBP antibody are specific for PKCαb and both PKCβ paralogs. PKCα NBP detects a faint band in all sam- ples, including mock controls, likely recognizing an endog- enous distantly related PKC orthologue present in HEK293T cells (asterisk at 75 kDa), PKCα MC5 recognizes both zebrafish PKCα paralogs (b, (1) and (2) lane) as well as PKCβb (b, 4. lane). The PKCβ1 antibody also recognizes PKCαa (c, (1) lane) but PKCβb only faintly (c, 4. lane), and PKCαb not at all (c, (2) lane). PKCγ lysates were not specifically detected with any antibody used (a–c, 5. lane). Anti-Vinculin (124 kDa) antibodies were used as loading controls. The different antibody recognition patterns are sum- marized in panel d

Article Snippet: Another PKCα antibody ([Novus (NBP)] also weakly labels the same cells in the INL but showed a very strong labeling in the retinal ONL, presumably in accessory outer segments (Fig. 1c′′-′′) (Hodel et al. 2014).

Techniques: Western Blot, Recombinant

Journal: Histochemistry and cell biology

Article Title: Differential expression of PKCα and -β in the zebrafish retina.

doi: 10.1007/s00418-018-1764-8

Figure Lengend Snippet: Fig. 3 Western blot analysis of recombinant zebrafish cPKC proteins using PKCα and -β antibodies. Three different anti- bodies were used to compare the specificity of each antibody with recombinant zebrafish PKCs. The PKCα NBP antibody are specific for PKCαb and both PKCβ paralogs. PKCα NBP detects a faint band in all sam- ples, including mock controls, likely recognizing an endog- enous distantly related PKC orthologue present in HEK293T cells (asterisk at 75 kDa), PKCα MC5 recognizes both zebrafish PKCα paralogs (b, (1) and (2) lane) as well as PKCβb (b, 4. lane). The PKCβ1 antibody also recognizes PKCαa (c, (1) lane) but PKCβb only faintly (c, 4. lane), and PKCαb not at all (c, (2) lane). PKCγ lysates were not specifically detected with any antibody used (a–c, 5. lane). Anti-Vinculin (124 kDa) antibodies were used as loading controls. The different antibody recognition patterns are sum- marized in panel d

Article Snippet: Different primary anti-PKCα and -β antibodies were used at the following concentrations: PKCα NBP1-19273, 1:500 (Novus, Abingdon, UK); PKCα MC5, NB 200–568, 1:1000 (Novus); PKCα MC5, 1:1000 (Genetex), PKCβ1 C16, SC209, 1:1000 (Santa Cruz Biotechnology, Heidelberg, Germany).

Techniques: Western Blot, Recombinant