Hybridoma (2005), 2010-10, Vol.29 (5), p.391-401
2010
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- Autor(en) / Beteiligte
- Titel
- Titin A-band-specific monoclonal antibody Tit1 5H1.1. Cellular Titin as a centriolar protein in non-muscle cells
- Ist Teil von
- Hybridoma (2005), 2010-10, Vol.29 (5), p.391-401
- Ort / Verlag
- United States
- Erscheinungsjahr
- 2010
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- We report the development of a new mouse anti-titin monoclonal antibody, named MAb Tit1 5H1.1, using the synthetic peptide corresponding to an amino acid sequence in the A-band of the titin molecule as immunogen. In the human skeletal muscle, MAb Tit1 5H1.1 reveals a clearly striated staining pattern, reacting with the A-band of the sarcomere. Electrophoretic, immunoblotting, and amino acid sequence analyses with ESI-MS/MS of human skeletal muscle tissue proved the target antigen of MAb Tit1 5H1.1 to be titin. The antibody reacts with titin also in non-muscle cells, producing a punctate pattern in cytoplasm and the nucleus. The most striking finding was a clear reaction of MAb Tit1 5H1.1 with centrioles in all cell types investigated so far. Immunocytochemical co-localization study with ninein-specific antibodies confirmed that the target antigen of MAb Tit1 5H1.1 is a centriole-associated protein. Experiments of the inhibition of synthesis of titin using titin siRNA duplex for the destruction of titin mRNA have shown a decreased staining of centrioles by MAb Tit1 5H1.1 in non-muscle cells and support the proposal that the target antigen of MAb is indeed titin. We suggest this anti-titin monoclonal antibody could be a valuable tool in the study of titin function and its subcellular location, both in muscle and non-muscle cells.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1554-0014eISSN: 1557-8348DOI: 10.1089/hyb.2009.0116Titel-ID: cdi_pubmed_primary_21050039
- Format
- –
- Schlagworte
- Amino Acid Sequence, Animals, Antibodies, Monoclonal - biosynthesis, Antibodies, Monoclonal - metabolism, Antibodies, Monoclonal - pharmacology, Antibody Specificity, Cells - drug effects, Cells - metabolism, Cells, Cultured, Centrioles - drug effects, Centrioles - metabolism, Connectin, Epitopes, Female, Humans, Immunoblotting, Mice, Mice, Inbred BALB C, Muscle Cells - drug effects, Muscle Cells - metabolism, Muscle Proteins - antagonists & inhibitors, Muscle Proteins - genetics, Muscle Proteins - immunology, Muscle Proteins - metabolism, Protein Kinases - genetics, Protein Kinases - immunology, Protein Kinases - metabolism, RNA, Small Interfering - pharmacology, Spectrometry, Mass, Electrospray Ionization