The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1.25 µg/ml. Predicted molecular weight: 57 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use a concentration of 4 - 8 µg/ml.
Use at an assay dependent concentration.
Titre using peptide based assay: 1/312500.
UPF3B is part of a post-splicing multiprotein complex involved in both mRNA nuclear export and mRNA surveillance. mRNA surveillance detects exported mRNAs with truncated open reading frames and initiates nonsense-mediated mRNA decay (NMD). When translation ends upstream from the last exon-exon junction, this triggers NMD to degrade mRNAs containing premature stop codons. UPF3B binds to the mRNA and remains bound after nuclear export, acting as a nucleocytoplasmic shuttling protein. It forms with Y14 a complex that binds specifically 20 nt upstream of exon-exon junctions. This gene is located on the long arm of chromosome X (referenced from entrez gene).
Nucleus. Cytoplasm. Note: Shuttling between the nucleus and the cytoplasm.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling UPF3B/RENT3B with ab66753 at 4-8µg/ml. Arrows indicate positively labeling epithelial cells of the fundic gland. Magnification: 400X.
Western blot - UPF3B/RENT3B antibody (ab66753)
Anti-UPF3B/RENT3B antibody (ab66753) at 1.25 µg/ml + Jurkat cell lysate at 10 µg
Secondary HRP-conjugated anti-rabbit IgG at 1/50000 dilution