EBP50 Antibody (PrecisionAbTM Antibody)

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EBP50 Antibody (PrecisionAb<sup>TM</sup> Antibody) gallery image 1

Western blot analysis of whole cell lysates probed with EBP50 antibody followed by detection with HRP conjugated Goat anti Mouse IgG (1/10,000, STAR207P) and visualized on the ChemiDoc MP with 11 second exposure. Arrow points to EBP50 (molecular weight 50 kDa)

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  • EBP50 Antibody with Control Lysate
  • Mouse anti EBP50
  • EBP50 Antibody with Control Lysate
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    EBP50 antibody recognizes Na(+)/H(+) exchange regulatory cofactor NHE-RF1, also known as NHERF-1, ezrin-radixin-moesin-binding phosphoprotein 50, EBP50, regulatory cofactor of Na(+)/H(+) exchanger, sodium-hydrogen exchanger regulatory factor 1 or solute carrier family 9 isoform A3 regulatory factor 1. The antibody has been extensively validated for western blotting using whole cell lysates.
    • Product Type
      PrecisionAb™ Monoclonal
    • Isotype
      IgG2b
    • Format
      Purified
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      VMA00054WBdatasheet pdfdatasheet pdf100 µl
      VMA00054
      VMA00054KWBdatasheet pdfdatasheet pdf datasheet pdf10 Westerns
      VMA00054K
      VMA00054KTWBdatasheet pdfdatasheet pdf datasheet pdf2 Westerns
      VMA00054KT
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      References
      Reviews
      -
      • Mouse anti Human EPB50 antibody recognizes ezrin/radixin/moesin (ERM)-binding phosphoprotein of 50 kDa (EBP50) otherwise known as Na+/H+ exchanger regulatory factor 1 (NHERF1), regulatory cofactor of Na(+)/H(+) exchanger, sodium-hydrogen exchanger regulatory factor 1 or solute carrier family 9 isoform A3 regulatory factor 1. EBP50 is a 358 amino acid cytoplasmic adaptor molecule containing two PDZ domains. EBP50 is expressed in tissues containing polarized epithelia, including the small intestine, placenta, liver and kidney.

        The ERM family of membrane-cytoskeletal linking proteins act to maintain a regulated linkage between actin filaments (F-actin) and the plasma membrane. Studies have revealed that EBP50 is an important ERM binding partner, which interacts with the N-terminal FERM (four point one ERM) domain of ERM proteins, via its C-terminal domain. EBP50 colocalizes with actin and ezrin, in the apical mircovilli of epithelial cells and this role as binding partner has been further shown through the interaction of EBP50 with the transmembrane adaptor protein known as protein associated with glycosphingolipid-enriched microdomains (PAG), in T-cells (Reczek et al. 1997).

        Mutations in the gene encoding EBP50 leads to nephrolithiasis/osteoporosis, hypophosphatemic, 2 disease, characterized by impaired phosphate metabolism (Courbebaisse et al. 2012). Other ligands which bind to the PDZ domains of EBP50, include beta-catenin, cystic fibrosis transmembrane conductance regulator and the type II Na/Pi cotransporter Npt2, implicating EBP50 in a diverse range of biological systems.

        Mouse anti Human EBP50 antibody recognizes EBP50 as a single band of ~50 kDa in western blotting in multiple cell line lysates including myeloid, lymphoid, neuronal, adenocarcinoma, sarcoma and embryonic kidney.
      • Target Species
        Human
      • Western Blotting
        Anti EBP50 antibody detects a band of approximately 50 kDa in Jurkat cell lysates.
        Anti EBP50 antibody detects a band of approximately 50 kDa in Jurkat cell lysates.
        Anti EBP50 antibody detects a band of approximately 50 kDa in Jurkat cell lysates.
      • Product Form
        Purified IgG - liquid
        Purified IgG - liquid
        Purified IgG - liquid
      • Preparation
        Pack Size: 10 Westerns100μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from tissue culture supernatant
        Pack Size: 2 Westerns20μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from tissue culture supernatant
        Mouse monoclonal antibody purified by affinity chromatography on Protein G from tissue culture supernatant
        Pack Size: 10 Westerns100μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from tissue culture supernatant
        Pack Size: 2 Westerns20μl Mouse monoclonal antibody purified by affinity chromatography on Protein G from tissue culture supernatant
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
        Phosphate buffered saline
      • Preservative Stabilisers
        0.09% Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
        0.09% Sodium Azide (NaN3)
      • Immunogen
        Bacterially produced recombinant full length human EBP50.
      • Instructions for Use
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
        Please refer to the PrecisionAb western blotting protocol. For additional information on secondary antibody dilution and exposure time see product web page.
      • Lysate Composition
        400μg Jurkat lysate lyophilized in RIPA buffer.
        400μg Jurkat lysate lyophilized in RIPA buffer.
      • Lysate Reconstitution
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
        - If using DDT reconstitute the lyophilized lysate with 190μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 10μl 2M DTT.
        - If using BME reconstitute the lyophilized lysate with 180μl DI H2O, add 200μl 2x Laemmli Sample Buffer and 20μl BME.
        Heat at 95°C for 5 minutes. For 10 well mini gels load 25μl. For other gel and comb formats please refer to the PrecisionAb western blotting protocol.
      • Storage
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
        Store undiluted at -20oC, avoiding repeated freeze thaw cycles.
        Antibody: Store undiluted at -20oC, avoiding repeated freeze thaw cycles.

        Lysate: Store lyophilized lysate at -20°C. After reconstitution aliquot and store at -20°C for up to 3 months or at -80°C for longer term storage.
      • Shelf Life
        As supplied, 12 months from date of despatch.
        As supplied, 12 months from date of despatch.
        As supplied, 12 months from date of despatch.
      • Acknowledgements
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
        PrecisionAb™ is a trademark of Bio-Rad Laboratories.
      • Regulatory
        For research purposes only
      • GO Terms
        microvillus
        intracellular membrane-bounded organelle
        apical plasma membrane
        centrosome
        protein complex assembly
        actin cytoskeleton
        Wnt receptor signaling pathway
        beta-2 adrenergic receptor binding
        endomembrane system
        filopodium
        ruffle
        regulation of sodium:hydrogen antiporter activity
        PDZ domain binding
      • UniProt
      • Entrez Gene
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000
      • Application NameYesNoMin DilutionMax Dilution
        Western Blotting1/1000

      • PrecisionAb antibodies have been extensively validated for the western blot application. The antibody has been validated at the suggested dilution. Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Further optimization may be required dependant on sample type.
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
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        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
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        Recommended Negative Isotype Control

          Useful Reagents

            Recommended Positive Controls

              Histology Controls

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