Contaminating reactivity of a monoclonal CCAAT/Enhancer Binding Protein β antibody in differentiating myoblasts
| dc.contributor.author | AlSudais, Hamood | |
| dc.contributor.author | Wiper-Bergeron, Nadine | |
| dc.date.accessioned | 2019-11-03T04:23:50Z | |
| dc.date.available | 2019-11-03T04:23:50Z | |
| dc.date.issued | 2019-10-31 | |
| dc.date.updated | 2019-11-03T04:23:50Z | |
| dc.description.abstract | Abstract Objective CCAAT/Enhancer Binding proteins (C/EBPs) are transcription factors involved in the regulation of a variety of cellular processes. We used the Abcam Recombinant Anti-C/EBP beta antibody (E299) to detect C/EBPβ expression during myogenesis. Though the antibody is monoclonal, and the immunogen used is highly specific to C/EBPβ, we identified an intense band at 23 kDa on western blot that did not correspond to any of the known isoforms of C/EBPβ, or family members predicted to cross-react. Absent in myoblast cells overexpressing C/EBPβ, the band was present when C/EBPβ was knocked down, confirming specificity for a protein other than C/EBPβ. The objective of this work was to identify the contaminating reactivity. Results We performed immunoprecipitation followed by mass spectrometry to identified myosin light chain 4 (MYL4) as the unknown band, suggesting that the Abcam monoclonal antibody directed against C/EBPβ is not pure, but contains a contaminating antibody against MYL4. Caution should be used when working in cells lines that express MYL4 to not confound the detection of MYL4 with that of C/EBPβ isoforms. | |
| dc.identifier.citation | BMC Research Notes. 2019 Oct 31;12(1):717 | |
| dc.identifier.uri | https://doi.org/10.1186/s13104-019-4749-3 | |
| dc.identifier.uri | https://doi.org/10.20381/ruor-24048 | |
| dc.identifier.uri | http://hdl.handle.net/10393/39805 | |
| dc.language.rfc3066 | en | |
| dc.rights.holder | The Author(s) | |
| dc.title | Contaminating reactivity of a monoclonal CCAAT/Enhancer Binding Protein β antibody in differentiating myoblasts | |
| dc.type | Journal Article |
