Generation and characterization of monoclonal antibodies to TDRD7 protein.
TDRD7 is a scaffold protein whose specific function is unknown. It has been identified in complexes with proteins that regulate cytoskeleton dynamics and centrosomal movements, mRNA transport, and protein translation apparatus. Here we report the generation and characterization of monoclonal antibodies against TDRD7 protein. Bacterially expressed His-tagged fragments of TDRD7 were used as antigens. Spleen cells from immunized mice were collected and fused with SP2/0 myeloma cells using PEG 2000. High titer anti-TDRD7 antibody-producing hybridoma cell lines were identified by enzyme-linked immunosorbent assay (ELISA) and then subcloned by limiting dilution. Antibodies produced by E6 clone were further tested for their reactivity with the TDRD7 recombinant proteins. The results obtained clearly indicate that E6 anti-TDRD7 antibodies recognize specifically recombinant 6His-tagged TDRD7 proteins and endogenous TDRD7 in Western blotting, immunoprecipitation, and immunocytochemistry. In summary, these antibodies will be useful for researchers investigating TDRD7 and molecular complexes involving this protein.
Skorokhod
Oleksandr
O
Department of Cell Signalling, Institute of Molecular Biology and Genetics of NAS of Ukraine, Kyiv, Ukraine.
Nemazanyy
Ivan
I
Breus
Oksana
O
Filonenko
Valeriy
V
Panasyuk
Ganna
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012261:Ribonucleoproteins / Q000737:chemistry*; D013154:Spleen / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; C517522:TDRD7 protein, mouse