Journal: Aquaculture

Article Title: Morpholino oligomer delivery via bath immersion for use in reverse genetic studies on the early development of eastern oysters (Crassostrea virginica)

doi: 10.1016/j.aquaculture.2025.742261

Figure Lengend Snippet: Fig. 3. Western blot analyses of the antiserums generated against eastern oyster Gcl, mammalian GMCL1, eastern oyster Vasa, and zebrafish Vasa. The molecular weight of eastern oyster Gcl and Vasa proteins were expected to be 73 kDa and 77–83 kDa, respectively, calculated from deduced amino acids. Total protein from oyster ovary and testis were first separated by SDS-PAGE and then allowed to bind (A) with three eastern oyster Gcl antibodies generated by Genescript and a commercial mammalian GMCL1 antibody (Proteintech 15,575–1-AP); (B) with three eastern oyster Vasa antibodies generated by Genescript; (C) with a polyclonal antibody against zebrafish Vasa (zfVasa, GeneTex GTX128306). None of the Genescript and Protrintech antibodies had high specificity for the target proteins since multiple bands were detected, while the zfVasa antibody cross-reacted and dectected oyster Vasa protein. The following dilutions were used for the primary anti bodies, 1:200 (Gcl-1, Gcl-2); 1:400 (Gcl-3); 1:1500 (GMCL1); 1:600 (Vasa-1); 1:100 (Vasa-2); 1:10,000 (Vasa-3); 1:10,000 (zfVasa). O, ovary; T, testis; M, protein marker. The numbers next to the bands in the protein marker lane indicate the molecular weights in kDa.

Article Snippet: Additionally, a polyclonal antibody against the mammalian Gcl homolog GMCL1 (Proteintech, 15,575–1-AP) and a polyclonal antibody against zebrafish Vasa (GeneTex, GTX128306) was also purchased for testing cross-reactivity with eastern oyster Gcl and Vasa proteins, respectively.

Techniques: Western Blot, Generated, Molecular Weight, SDS Page, Marker