The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use at an assay dependent concentration.
1/2000 - 1/10000. Detects a band of approximately 117 kDa (predicted molecular weight: 109 kDa).
Use at 2-5 µg/mg of lysate.
Component of the U5 snRNP complex required for pre-mRNA splicing. Binds GTP.
Involvement in disease
Defects in EFTUD2 are the cause of mandibulofacial dysostosis with microcephaly (MFDM) [MIM:610536]. A rare syndrome characterized by progressive microcephaly, midface and malar hypoplasia, micrognathia, microtia, dysplastic ears, preauricular skin tags, significant developmental delay, and speech delay. Many patients have major sequelae, including choanal atresia that results in respiratory difficulties, conductive hearing loss, and cleft palate.
Belongs to the GTP-binding elongation factor family. EF-G/EF-2 subfamily.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma (left) and mouse teratoma (right) tissues labelling EFTUD2 with ab72456 at 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-EFTUD2 antibody (ab72456)
All lanes : Anti-EFTUD2 antibody (ab72456) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
HeLa whole cell lysate (1 mg for IP, 20% of IP loaded), with ab72456 used for IP at 3 µg/mg lysate, and at 0.1 µg/ml for subsequent WB. Detection: Chemiluminescence with an exposure time of 10 seconds.
IHC image of ab72456 staining in human melanoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72456, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunocytochemistry/ Immunofluorescence - Anti-EFTUD2 antibody (ab72456)This image is courtesy of an anonymous Abreview
ab110240 staining EFTUD2 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence).
Cells were fixed with formaldehyde, permeabilized with 0.3% Triton X-100 and blocked with 5% donkey serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/1000 in PBS) for 1.5 hours at 25°C. A Cy3®-conjugated sheep anti-rabbit polyclonal IgG (1/100) was used as the secondary antibody. DNA was stained with TO-PRO®-3.
Cloutier P et al. R2TP/Prefoldin-like component RUVBL1/RUVBL2 directly interacts with ZNHIT2 to regulate assembly of U5 small nuclear ribonucleoprotein. Nat Commun8:15615 (2017).
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