Recombinant Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5562] to EBP50/NHERF-1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-EBP50/NHERF-1 antibody [EPR5562]
See all EBP50/NHERF-1 primary antibodies -
Description
Rabbit monoclonal [EPR5562] to EBP50/NHERF-1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HCT116, HepG2, 293T, Jurkat, C6, PC-12 and MCF-7 cell lysates; Human kidney tissue
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5562 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109430 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 |
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WB |
1/1000 - 1/10000. Predicted molecular weight: 39 kDa.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 39 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 9368 Human
- Entrez Gene: 59114 Rat
- Omim: 604990 Human
- SwissProt: O14745 Human
- SwissProt: Q9JJ19 Rat
- Unigene: 724482 Human
- Unigene: 728760 Human
- Unigene: 35142 Rat
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Alternative names
- EBP 50 antibody
- EBP50 antibody
- Ezrin radixin moesin binding phosphoprotein 50 antibody
see all
Images
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All lanes : Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SLC9A3R1 knockout HeLa cell lysate
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 46 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-EBP50/NHERF-1 antibody [EPR5562] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109430 was shown to bind specifically to EBP50/NHERF-1. A band was observed at 46 kDa in wild-type HeLa cell lysates with no signal observed at this size in SLC9A3R1 knockout cell line ab264914 (knockout cell lysate ab257280). To generate this image, wild-type and SLC9A3R1 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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All lanes : Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430) at 1/1000 dilution
Lane 1 : Wild-type HCT116 cell lysate
Lane 2 : SLC9A3R1 knockout HCT116 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 39 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab109430 observed at 48 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109430 was shown to react with EBP50/NHERF-1 in wild-type HCT116 cells in western blot. Loss of signal was observed when knockout cell line ab266876 (knockout cell lysate ab257281) was used. Wild-type HCT116 and SLC9A3R1 knockout HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109430 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemical analysis of EBP50/NHERF-1 in paraffin-embedded Human kidney tissue using ab109430 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: EBP50/NHERF-1 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109430 observed at 48 kDa. Red - loading control, ab18058, observed at 124 kDa.
ab109430 was shown to specifically recognize EBP50/NHERF-1 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when EBP50/NHERF-1 knockout samples were usexamined. Wild-type and EBP50 knockout samples were subjected to SDS-PAGE. ab109430 and ab18058 (loading control to Vinculin) were diluted at 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
All lanes : Anti-EBP50/NHERF-1 antibody [EPR5562] (ab109430) at 1/1000 dilution
Lane 1 : HepG2 cell lysate
Lane 2 : 293T cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : C6 cell lysate
Lane 5 : PC12 cell lysate
Lane 6 : MCF7 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 39 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab109430 has been referenced in 2 publications.
- Sikorski K et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371
- Feng D et al. Reduced EBP50 expression levels are correlated with unfavorable clinicopathological features of extrahepatic bile duct carcinoma and promote the proliferation and migration of QBC939 cells. Oncol Lett 13:2758-2764 (2017). IHC-P ; Human . PubMed: 28454463