|Detection of Human Phospho-eIF4B (S422) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes or 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-eIF4B (S422) Monoclonal Antibody (Catalog # MAB3800) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-eIF4B (S422) at approximately 93 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
eIF4B (eukaryotic translation initiation factor 4B), an RNA binding protein, is essential for the binding of mRNA to the 43S pre-initiation complex, which consists of the 40S ribosomal subunit bound to a ternary complex of eIF2, GTP, and Met-tRNA stabilized by eIF3. An arginine rich motif (ARM) in the carboxy-terminus of eIF4B binds RNA non-specifically, while a canonical RNA motif (RMM) near the amino terminus binds specifically to 18S rRNA. The simultaneous binding of specific and nonspecific RNA may serve to facilitate the binding of the 40S subunit to the mRNA by serving as a bridge between the 18S rRNA and the mRNA. Besides its binding activity, eIF4B in conjunction with the heterotrimeric eIF4F, stimulates the ATPase and RNA helicase activity of eIF4E.
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