Overview

  • Product name

    Anti-EBP50/NHERF-1 antibody
    See all EBP50/NHERF-1 primary antibodies
  • Description

    Rabbit polyclonal to EBP50/NHERF-1
  • Host species

    Rabbit
  • Specificity

    Detects EBP50/NHERF-1 from human T84 and Calu3 cells as well as recombinant human protein.
  • Tested applications

    Suitable for: ICC/IF, IHC-Fr, IP, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Dog, Human
    Predicted to work with: Rabbit, Cow, Cynomolgus monkey
  • Immunogen

    Recombinant fragment (His-tag) corresponding to Human EBP50/NHERF-1. All His-tag specific antibody has been removed.
    Sequence:

    RSASSDTSEELNSQDSP

  • Positive control

    • T84 cell extract, mouse airway epithelium

Properties

Applications

Our Abpromise guarantee covers the use of ab3452 in the following tested applications.

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

Application Abreviews Notes
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr Use at an assay dependent concentration. PubMed: 15703401
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 50 kDa).
IHC-P Use a concentration of 10 µg/ml.

Target

  • Function

    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.
  • Tissue specificity

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

    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).
  • Sequence similarities

    Contains 2 PDZ (DHR) domains.
  • Post-translational
    modifications

    Phosphorylated on serine residues.
  • Cellular localization

    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
  • Database links

  • Alternative names

    • 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

Images

  • All lanes : Anti-EBP50/NHERF-1 antibody (ab3452) at 1/1000 dilution

    All lanes : Whole cell lysate prepared from human HepG2 cells

    Secondary
    All lanes : HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa


    Exposure time: 40 seconds

    See Abreview

  • All lanes : Anti-EBP50/NHERF-1 antibody (ab3452) at 1/1000 dilution

    All lanes : Whole tissue lysate prepared from mouse liver

    Secondary
    All lanes : HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 50 kDa


    Exposure time: 30 seconds

    See Abreview

  • Western blot detection of recombinant Human EBP50/NHERF-1 using ab3452.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse airway tissue sections labeling EBP50/NHERF-1 with ab3452.

References

This product has been referenced in:

  • Tsurudome Y  et al. Circadian clock component PERIOD2 regulates diurnal expression of Na+/H+ exchanger regulatory factor-1 and its scaffolding function. Sci Rep 8:9072 (2018). Read more (PubMed: 29899468) »
  • Henkel B  et al. NHERF1 in Microvilli of Vomeronasal Sensory Neurons. Chem Senses 42:25-35 (2017). Read more (PubMed: 27655939) »
See all 30 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (hepatocytes)
Permeabilization
Yes - 0.1% Triton X-100
Specification
hepatocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 17 2014

Application
Western blot
Sample
Cow Tissue lysate - whole (Kidney tissue lysate)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
20 µg
Specification
Kidney tissue lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 10 2011

Application
Western blot
Sample
Human Cell lysate - whole cell (HepG2)
Loading amount
20 µg
Specification
HepG2
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Oct 30 2008

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Loading amount
25 µg
Specification
liver
Gel Running Conditions
Reduced Denaturing (8)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Oct 30 2008

Answer

Thank you for your patience in this matter. I have received a reply from the originator of ab3452 (EBP50 antibody). The epitope recognized by this product is amino acids 286-302 of the human EBP50 protein. This information has been added to the product datasheet. I hope this helps with the reviewers. Good luck with your manuscript submission. Please don't hesitate to contact us again if you have further questions.

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 70213 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE human cell lines HL60, Jurket and BJAB. whole cell lysates PRIMARY ANTIBODY ab3452: used at 1:1000, incubate at room temperature for 1 hour, wash 5 times and 15 minutes per wash ab9526: as above SECONDARY ANTIBODY ab3452: Goat anti rabbit IgG, 1:5000, [a competitor], incubated and washed per 1' ab9526: sheep anti mouse IgG, 1:5000, [a competitor], incubated and washed per 1' DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED ab3452: rabbit polyclonal anti-moesin [a competitor] ab9526: KPL1 which is a mouse monoclonal (IgG) to human PSGL-1 Both negative controls always gave us a band of the proper MW ANTIBODY STORAGE CONDITIONS The antibody was aliquoted and is stored at -20C. Working stock is thawed on ice and stored a 4C. SAMPLE PREPARATION 1% TX-100 lysis buffer with 4 different protease inhibitors. Samples were boiled for 2 minutes in SDS sample buffer AMOUNT OF PROTEIN LOADED WCL from 4 x 10^6 cells ELECTROPHORESIS/GEL CONDITIONS Non reduced, 7.5 % separating and 4 % stacking TRANSFER AND BLOCKING CONDITIONS Standard western transfer buffer, 1 hour, blocked with 2 % gelatin for 2 hours. MW markers transfered to Nitrocellulose (could visualize these as intense bands!) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES The comments are for both Abs I purchased from abcam. This is the second comment form I have filled out, I believe the first one was emailed on4/6/05. ab3452: gave us a very weak signal of the appropriate MW, no background bands were noted. ab9526: no appropriate signal and no background

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Answer

I'm sorry to hear you are having problems with ab3452 and ab9526 and you had to re-send us your protocol details, I appreciate you taking the time to let us know those. I would like to recommend incubating the primary antibodies overnight at 4C and to try more concentrated antibody (e.g 1:200-1:500). A good positive control for EBP50 is T84 cell lysate. Please do not hesitate to contact us again if you still have problems with the positive control and those recommendations,

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