| First Author | Hunt R | Year | 2007 |
| Journal | Hybridoma (Larchmt) | Volume | 26 |
| Issue | 4 | Pages | 231-40 |
| PubMed ID | 17725385 | Mgi Jnum | J:127531 |
| Mgi Id | MGI:3763857 | Doi | 10.1089/hyb.2007.0507 |
| Citation | Hunt R, et al. (2007) Generation and characterization of mouse monoclonal antibody 5E1 against human transcription factor GLI3. Hybridoma (Larchmt) 26(4):231-40 |
| abstractText | GLI3 is a transcriptional effector of the developmentally important hedgehog (Hh) signaling pathway. Here we report the production of mouse monoclonal antibody (MAb) against putative repressive motif in GLI3 (GLI3pRM). BALB/c mice were immunized with purified recombinant human GLI3pRM protein, and the splenocytes from these mice were fused with myeloma cell line (SP2/0) by using standard hybridoma production techniques. Resulting hybridomas producing anti-GLI3pRM antibodies were screened by enzyme-linked immunosorbent assay (ELISA) and isotyped. The specificity of MAb 5E1 was determined based on its activity in Western blot and immunofluorescence analyses of the human NT2/D1 cell line. The results showed that MAb 5E1 was immunoglobulin IgM/kappa, recognizing recombinant human GLI3pRM specifically. In addition, MAb 5E1 bound to the full-length (FL-GLI3) as well as a short protein (GLI3R) and did not cross-react with a similar region in GLI2. MAb 5E1 could also be used to detect the expression of Gli3 in mouse cell lines and embryonic tissues. |
