Anti-EBP50/NHERF-1 抗体 [EBP-10] (ab9526)

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Key features and details

  • Mouse monoclonal [EBP-10] to EBP50/NHERF-1
  • Suitable for: Flow Cyt, WB
  • Reacts with: Human
  • Isotype: IgG2b

製品の概要

  • 製品名

    Anti-EBP50/NHERF-1 antibody [EBP-10]
    EBP50/NHERF-1 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [EBP-10] to EBP50/NHERF-1

  • 由来種

    Mouse

  • 特異性

    Human ERM (ezrin/radixin/moesin)-binding phosphoprotein of 50 kDa (EBP50) / Na+/H+ exchange regulatory factor (NHERF-1)

  • アプリケーション

    適用あり: Flow Cyt, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant full length protein corresponding to Human EBP50/NHERF-1.

  • ポジティブ･コントロール

    • RAJI human lymphoma cell lysate.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab9526 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
WB Use at an assay dependent concentration. Recommended dilution: 2µg/ml. Sample preparation: Lysis buffer with maltoside (1%). Application note: Non-reducing conditions with SDS-PAGE (10% separating gel).

ターゲット情報

  • 機能

    Scaffold protein that connects plasma membrane proteins with members of the ezrin/moesin/radixin family and thereby helps to link them to the actin cytoskeleton and to regulate their surface expression. Necessary for recycling of internalized ADRB2. Was first known to play a role in the regulation of the activity and subcellular location of SLC9A3. Necessary for cAMP-mediated phosphorylation and inhibition of SLC9A3. May enhance Wnt signaling. May participate in HTR4 targeting to microvilli (By similarity). Interacts with MCC.

  • 組織特異性

    Detected in liver, kidney, pancreas, prostate, spleen, small intestine and placenta, in particular in the syncytiotrophoblast.

  • 関連疾患

    Defects in SLC9A3R1 are the cause of hypophosphatemic nephrolithiasis/osteoporosis type 2 (NPHLOP2) [MIM:612287]. Hypophosphatemia results from idiopathic renal phosphate loss. It contributes to the pathogenesis of hypophosphatemic urolithiasis (formation of urinary calculi) as well to that of hypophosphatemic osteoporosis (bone demineralization).

  • 配列類似性

    Contains 2 PDZ (DHR) domains.

  • 翻訳後修飾

    Phosphorylated on serine residues.

  • 細胞内局在

    Cytoplasm. Apical cell membrane. Endomembrane system. Cell projection > filopodium. Cell projection > ruffle. Cell projection > microvillus. Translocates from the cytoplasm to the apical cell membrane in a PODXL-dependent manner (By similarity). Colocalizes with actin in microvilli-rich apical regions of the syncytiotrophoblast. Found in microvilli, ruffling membrane and filopodia of HeLa cells. Present in lipid rafts of T-cells.
  • Information by UniProt

  • 参照データベース

    • 別名

      • EBP 50 antibody
      • EBP50 antibody
      • Ezrin radixin moesin binding phosphoprotein 50 antibody
      • Ezrin-radixin-moesin-binding phosphoprotein 50 antibody
      • Na(+)/H(+) exchange regulatory cofactor NHE RF antibody
      • Na(+)/H(+) exchange regulatory cofactor NHE-RF1 antibody
      • Na+/H+ exchange regulatory co factor antibody
      • NHERF 1 antibody
      • NHERF antibody
      • NHERF-1 antibody
      • NHERF1 antibody
      • NHRF1_HUMAN antibody
      • NPHLOP2 antibody
      • Regulatory cofactor of Na(+)/H(+) exchanger antibody
      • Slc9a3r1 antibody
      • Sodium hydrogen exchanger regulatory factor 1 antibody
      • Sodium-hydrogen exchanger regulatory factor 1 antibody
      • Sodium/hydrogen exchanger regulatory factor 1 antibody
      • Solute carrier family 9 (sodium/hydrogen exchanger) member 3 regulator 1 antibody
      • Solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulatory factor 1 antibody
      • Solute carrier family 9 isoform 3 regulatory factor 1 antibody
      • Solute carrier family 9 isoform A3 regulatory factor 1 antibody
      • Solute carrier family 9 member 3 regulator 1 antibody
      see all

    画像

    • Western blot - Anti-EBP50/NHERF-1 antibody [EBP-10] (ab9526)
      Western blot - Anti-EBP50/NHERF-1 antibody [EBP-10] (ab9526)
      All lanes : Anti-EBP50/NHERF-1 antibody [EBP-10] (ab9526) at 2 µg/ml

      Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
      Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Observed band size: 48 kDa
      why is the actual band size different from the predicted?

    • Flow Cytometry - Anti-EBP50/NHERF-1 antibody [EBP-10] (ab9526)
      Flow Cytometry - Anti-EBP50/NHERF-1 antibody [EBP-10] (ab9526)
      Overlay histogram showing HepG2 cells stained with ab9526 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9526, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    参考文献 (10)

    ab9526 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab9526 は 10 報の論文で使用されています。

    • He P  et al. The NHERF1 PDZ1 domain and IRBIT interact and mediate the activation of Na+/H+ exchanger 3 by ANG II. Am J Physiol Renal Physiol 311:F343-51 (2016). PubMed: 27279487
    • He P  et al. Restoration of Na+/H+ exchanger NHE3-containing macrocomplexes ameliorates diabetes-associated fluid loss. J Clin Invest 125:3519-31 (2015). PubMed: 26258413
    • Liu H  et al. SLC9A3R1 stimulates autophagy via BECN1 stabilization in breast cancer cells. Autophagy 11:2323-34 (2015). PubMed: 26218645
    • Wang B  et al. Ezrin-anchored PKA Coordinates Phosphorylation-dependent Disassembly of a NHERF1 Ternary Complex to Regulate Hormone-sensitive Phosphate Transport. J Biol Chem : (2012). PubMed: 22628548
    • Liu L  et al. Na+/H+ exchanger regulatory factor 1 (NHERF1) directly regulates osteogenesis. J Biol Chem 287:43312-21 (2012). WB ; Mouse . PubMed: 23109343
    • Reining SC  et al. GABARAP deficiency modulates expression of NaPi-IIa in renal brush-border membranes. Am J Physiol Renal Physiol 296:F1118-28 (2009). PubMed: 19225049
    • Saotome I  et al. Ezrin is essential for epithelial organization and villus morphogenesis in the developing intestine. Dev Cell 6:855-64 (2004). PubMed: 15177033
    • Helms C  et al. A putative RUNX1 binding site variant between SLC9A3R1 and NAT9 is associated with susceptibility to psoriasis. Nat Genet 35:349-56 (2003). PubMed: 14608357
    • Mahon MJ  et al. Na+/H+ exchanger-regulatory factor 1 mediates inhibition of phosphate transport by parathyroid hormone and second messengers by acting at multiple sites in opossum kidney cells. Mol Endocrinol 17:2355-64 (2003). PubMed: 12881509
    • Brdickova N  et al. Interaction between two adapter proteins, PAG and EBP50: a possible link between membrane rafts and actin cytoskeleton. FEBS Lett 507:133-6 (2001). PubMed: 11684085

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    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE whole cell lysates from human and mouse in SDS Sample buffer PRIMARY ANTIBODY AB9526 (1:1000) DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED mouse (-) human (+) ANTIBODY STORAGE CONDITIONS used within one week of receipt. maintained at 4oC. SAMPLE PREPARATION SDS sample buffer with 2-ME. heated 3 min at 95oC. Chilled/spun/loaded AMOUNT OF PROTEIN LOADED 40, 20, 10 micrograms ELECTROPHORESIS/GEL CONDITIONS SDS Denaturing PAGE TRANSFER AND BLOCKING CONDITIONS standard transfer methods. companion gels with same samples transfered at same time in same apparatus then blotted with two other primary antibodies that work well. This controls for the effectiveness of the transfer procedure and also the secondary antibodies and ECL procedures SECONDARY ANTIBODY gt anti mouse-HRP (1:10,000) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? increased primary from 1:2000

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    Answer

    Thank you for your patience. I am sorry to hear that you are experiencing difficulties with this product ab9526 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I sincerely apologize that this product did not work well for you. It seems that there were some crucial information not included in the datasheet, information and that would otherwise help you obtain results in your experiment. I just received confirmation from the originator of the product and they recommend diluting the antibody at 2 ug/ml. Also, for this particular antibody, a non-reducing condition should be used, as this antibody recognizes protein in its non-reduced form. Therefore, beta-mercaptoehanol must be removed from the loading buffer. Other information that I received are suitable positive control (RAJI human lymphoma cell lysate), sample preparation (Lysis buffer with maltoside 1%), and SDS-PAGE 10% separating gel. Please rest assured that the datasheet has already been updated to show the changes. If it is possible to repeat the experiments, then please try using a non-reducing condition. However, since it is a datasheet mistake on our part, I would therefore be more than happy to offer you a free replacement vial of your choice (ab3452). Please let me know how you would like to proceed with your enquiry. Also, please inform me of your purchase details (purchase order number, contact information, shipping address/purchasing agent), so that I can immediately arrange for a replacement for you. I look forward to hearing from you again in order to resolve the matter. Have a nice day.

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    Question

    Could you please let me know the exact epitope against which the ab9526 antibody was raised?.

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    Answer

    The epitope for this antibody has not been determined.

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    Question

    I would like to use this antibody to detect the homologous dog protein. Is it known which epitope is recognized by the antibody ?

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    Answer

    All the information we have on species cross reactivity is specified on the datasheet. To our knowledge, cross reactivity with dog has not yet been tested and the epitope for this antibody has not been fully characterised. Should you decide to go ahead and purchase this product, please let us know how you get on and in return we will forward a reward of your choice, typically an Amazon gift voucher.

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    Question

    I can't access the given reference (I would like to!) but would like to know what dilution the antibody is used at for western blotting and immunoprecipitation please

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    Answer

    Try this antibody at 1:1000 for WB and adjust accordingly from the results obtained. For IP, please follow the guidelines in the IP protocol (linked on the right hand side of the Abcam homepage). The references are not accessible from the on-line datasheet. Please copy and paste them into a Medline search.

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