The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 69 kDa).
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Use a concentration of 1 µg/ml.
功能Probably involved in connections of major cytoskeletal structures to the plasma membrane. In epithelial cells, required for the formation of microvilli and membrane ruffles on the apical pole. Along with PLEKHG6, required for normal macropinocytosis.
组织特异性Expressed in cerebral cortex, basal ganglia, hippocampus, hypophysis, and optic nerve. Weakly expressed in brain stem and diencephalon. Stronger expression was detected in gray matter of frontal lobe compared to white matter (at protein level). Component of the microvilli of intestinal epithelial cells. Preferentially expressed in astrocytes of hippocampus, frontal cortex, thalamus, parahippocampal cortex, amygdala, insula, and corpus callosum. Not detected in neurons in most tissues studied.
序列相似性Contains 1 FERM domain.
发展阶段Very strong staining is detected in the Purkinje cell layer and in part of the molecular layer of the infant brain compared to adult brain.
翻译后修饰Phosphorylated by tyrosine-protein kinases.
细胞定位Apical cell membrane. Cell projection. Cell projection > microvillus membrane. Cell projection > ruffle membrane. Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. Localization to the apical membrane of parietal cells depends on the interaction with MPP5. Localizes to cell extensions and peripheral processes of astrocytes (By similarity). Microvillar peripheral membrane protein.
Immunocytochemistry/ Immunofluorescence - Anti-Ezrin antibody (ab41672)This image is courtesy of an anonymous Abreview.
ab41672 staining Ezrin in undifferentiated human embryonic stem cells on a murine embryonic fibroblast (MEFs) feeding layer by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized and blocked with 1% serum for 30 minutes at room temperature. Samples were incubated with primary antibody (1/250 in 1% serum, 0.1% triton, 0.1% BSA + PBS) for 16 hours at 4°C. An Alexa Flour® 488-conjugated donkey anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/500.
Western blot - Ezrin antibody (ab41672)
All lanes : Anti-Ezrin antibody (ab41672) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 7 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Ezrin was immunoprecipitated using 0.5mg A431 whole cell extract, 5µg of Rabbit polyclonal Ezrin and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, A431 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab41672. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to anti-Rabbit HRP (IgG light chain) (ab99697). Band: 80kDa: Ezrin
IHC image of Ezrin staining in human breast carcinoma FFPE section, performed on a 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 ab41672, 1µg/ml, for 8 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.
Perisic L et al. Schip1 is a novel podocyte foot process protein that mediates actin cytoskeleton rearrangements and forms a complex with Nherf2 and ezrin. PLoS One10:e0122067 (2015).
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