RNA-binding protein that binds to iron-responsive elements (IRES), which are stem-loop structures found in the 5'-UTR of ferritin, and delta aminolevulinic acid synthase mRNAs, and in the 3'-UTR of transferrin receptor mRNA. Binding to the IRE element in ferritin results in the repression of its mRNA translation. Binding of the protein to the transferrin receptor mRNA inhibits the degradation of this otherwise rapidly degraded mRNA.
Background References
1. Samaniego F. et. al. Molecular characterization of a second iron-responsive element binding protein, iron regulatory protein 2. Structure, function, and post-translational regulation. J. Biol. Chem. 269:30904-30910(1994).
Western blot analysis of IREB2 on different lysates with Mouse anti-IREB2 antibody (HA600015) at 1/2,000 dilution.
Lane 1: Jurkat cell lysate (20 µg/Lane) Lane 2: A549 cell lysate (20 µg/Lane) Lane 3: Mouse liver tissue lysate (40 µg/Lane) Lane 4: Mouse kidney tissue lysate (40 µg/Lane) Lane 5: Rat liver tissue lysate (40 µg/Lane)
Predicted band size: 105 kDa Observed band size: 105 kDa
Exposure time: 11 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA600015) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP, AlpSdAbs® VHH(HRP) (001-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-IREB2 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 primary antibody (HA600015, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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