抗IRP2抗体 | Anti-IRP2 Antibody

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	Biological Strategies Validation. Western Blot: IRP2 Antibody [NB100-1798] - Raw macrophages (mouse) were either left untreated (lane 1) or treated overnight with 100mM DFO (lane 2) or 100mM hemin (lane 3). Proteins extracts were subjected to western blotting using IRP2 or b-actin antibodies diluted 1:500 in TBS-Tween.
	Immunohistochemistry-Paraffin: IRP2 Antibody [NB100-1798] - IHC analysis of a formalin fixed paraffin-embedded (FFPE) human placenta using 1:300 conc. of IRP2 antibody on a Bond Rx autostainer (Leica Biosystems). The assay involved 30 minutes of heat induced antigen retrieval (HIER) using 10mM sodium citrate buffer (pH 9.0) and endogenous peroxidase quenching with peroxide block. The sections were incubated with primary antibody for 15 minutes and Bond Polymer Refine Detection (Leica Biosystems) with DAB was used for signal development followed by counterstaining with hematoxylin. Cytoplasmic staining was observed in villi of endothelial cells.
	Western Blot: IRP2 Antibody [NB100-1798] - Detection of IRP2 in mouse liver lysate.
	Western Blot: IRP2 Antibody [NB100-1798] - Detection of IRP2 in RAW 264.7 lysate using NB100-1798. Lane 1: Lot B Lane 2: Lot C

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Background

Iron regulatory proteins -1 and -2 (IRP1, IRP2) are homologous cytoplasmic RNA-binding proteins that controls cellular iron metabolism and maintain systemic iron homeostasis in vertebrates along with serving some additional iron-independent functions. IRP2 belongs to aconitase/IPM isomerase family and it binds to IRES (iron-responsive elements), which are stem-loop structures found in 5'-UTR of ferritin, and delta aminolevulinic acid synthase mRNAs, and in the 3'-UTR of transferrin receptor mRNA. IRP2-IRE binding results in the repression of its mRNA translation and its binding to transferrin receptor mRNA inhibits the degradation of this otherwise quickly degradable mRNA. IRP2 is a substrate of FBXL5, an F-box protein that assembles together with Skp1, Cul1 and Rbx1 into an E3 ubiquitin ligase complex. When iron/O2 are high, IRP2 gets ubiquitinated/degraded by SCF complex containing FBXL5, whereas under conditions of low iron/O2, FBXL5 is degraded preventing IRP2 ubiquitination leading to its RNA-binding activity. IRP2 deficient mice depicts microcytosis, misregulated iron metabolism (in CNS, intestine as well as liver associated with FTH1 and FTL up-regulation) and iron deficiency in splenic macrophages associated with down-regulation of FTH1, FTL, and FPN1.

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