Spliceosomes are multi-protein complexes that are composed of snRNPs (small nuclear ribonucleoproteins) and a variety of associated protein factors, all of which work in concert to regulate the splicing of pre-mRNA. Snrp116, also known as EFTUD2 (elongation factor Tu GTP binding domain containing 2) or Snu114, is a 972 amino acid protein that localizes to the nucleus and belongs to the GTP-binding elongation factor family. Existing as a component of the multi-protein U5 snRNP spliceosome complex, Snrp116 plays an important role in pre-mRNA splicing, as well as in the recycling of spliceosomal snRNPs. The gene encoding Snrp116 maps to human chromosome 17, which comprises over 2.5% of the human genome and encodes over 1,200 genes.
Background References
1. Fabrizio P et al. An evolutionarily conserved U5 snRNP-specific protein is a GTP-binding factor closely related to the ribosomal translocase EF-2. EMBO J 16 (13): 4092–106 (1997).
2. Ewing et al. Large-scale mapping of human protein-protein interactions by mass spectrometry. Mol Syst Biol (England) 3 (1): 89 (2007).
Sequence Similarity
Belongs to the TRAFAC class translation factor GTPase superfamily. Classic translation factor GTPase family. EF-G/EF-2 subfamily.
Western blot analysis of EFTUD2 on different lysates with Rabbit anti-EFTUD2 antibody (R1411-3) at 1/1,000 dilution.
Lane 1: Jurkat cell lysate Lane 2: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane. Exposure time: 60 seconds;
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: R1411-3, 1/1,000 in 5% NFDM/TBST, room temperature for 2 hours Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 109 kDa Observed band size: 109 kDa
ICC staining EFTUD2 in B16F2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1411-3) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ICC staining EFTUD2 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1411-3) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
ICC staining EFTUD2 in SW480 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the antibody (R1411-3) at a dilution of 1:100 for 1 hour at room temperature, washed with PBS. Alexa Fluor™ 488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue using anti-EFTUD2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1411-3) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-EFTUD2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (R1411-3) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
Flow cytometric analysis of EFTUD2 was done on Hela cells. The cells were fixed, permeabilized and stained with EFTUD2 antibody at 1/100 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). After incubation of the primary antibody on room temperature for an hour, the cells was stained with a Alexa Fluor™ 488-conjugated goat anti-rabbit IgG Secondary antibody at 1/500 dilution.
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