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Immunofluorescent analysis of EBP50 was performed using 70% confluent log phase T-47D cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with EBP50 Rabbit Polyclonal Antibody (PA1-090) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Dog, Rat, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Bacterially expressed, His-tagged human EBP50.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||10 µg/ml|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-090 detects ezrin-radixin-moesin (ERM) binding phosphoprotein of 50 kDa (EBP50) from mouse tissues, human T84 and Calu3 cells as well as recombinant human protein.
PA1-090 has been successfully used in Western blot, Immunohistochemistry, immunoflourescence and immunoprecipitation procedures. By Western blot, this antibody detects an ~50 kDa protein representing EBP50 from T84 cell extract. Immunohistochemical staining of EBP50 in mouse airway results in staining of the epithelium.
The PA1-090 immunogen is recombinant human His-tagged EBP50 obtained from bacteria. All His-tag specific antibody has been removed. This product was epitope affinity purified using a synthetic peptide to residues C R(286) S A S S D T S E E L N S Q D S P(302) of human EBP50
EBP50 [ERM (ezrin-radixin-moesin) binding phosphoprotein of 50 kDa] is a PDZ containing protein that is involved in the linkage of integral membrane proteins to the cytoskeleton. EBP50 contains two tandem PDZ domains followed by a carboxy-terminal sequence that binds to members of the ERM family of membrane-cytoskeleton adaptors. The PDZ domains within EBP50 bind to the carboxy termini of such target proteins as beta2 adrenergic receptor, platelet-derived growth factor receptor (PDGFR), and the cystic fibrosis conductance regulator (CFTR). The PDZ domains are also believed to be involved in the oligomerization of EBP50 to form multiprotein complexes which helps to facilitate the formation of functional signalling complexes.
EBP50 has been shown to link integral membrane proteins to the cytoskeleton by binding to members of the ERM family of proteins, which bind to F-actin. Through these interactions, EBP50 is implicated in the localization of interactive groups of proteins into subcellular domains and in the regulation of activity of those interacting proteins. Immunohistochemical studies have shown that EBP50 is found exclusively at the apical membranes of epithelial cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Fine Needle Aspiration Cytology: A Tool to Study NHERF1 Expression as a Potential Marker of Aggressiveness in Lung Cancer.
PA1-090 was used in immunohistochemistry - paraffin section to assess the role of NHERF1 in lung tumors using fine needle aspirates
|Mangia A,Partipilo G,Schirosi L,Saponaro C,Galetta D,Catino A,Scattone A,Simone G||Molecular biotechnology (57:549)||2015|
Nuclear NHERF1 expression as a prognostic marker in breast cancer.
PA1-090 was used in immunohistochemistry to study the prognostic value of nuclear expression of NHERF-1 in breast cancer
|Paradiso A,Scarpi E,Malfettone A,Addati T,Giotta F,Simone G,Amadori D,Mangia A||Cell death & disease (4:null)||2013|
Characterization of CFTR High Expresser cells in the intestine.
PA1-090 was used in immunohistochemistry to study the function of CFTR High Expresser cells in the intestine of rats
|Jakab RL,Collaco AM,Ameen NA||American journal of physiology. Gastrointestinal and liver physiology (305:G453)||2013|
High density of tryptase-positive mast cells in human colorectal cancer: a poor prognostic factor related to protease-activated receptor 2 expression.
PA1-090 was used in immunohistochemistry to study the prognostic significance of a high density of tryptase-positive mast cells and PAR-2 expression levels in colorectal cancer patients
|Malfettone A,Silvestris N,Saponaro C,Ranieri G,Russo A,Caruso S,Popescu O,Simone G,Paradiso A,Mangia A||Journal of cellular and molecular medicine (17:1025)||2013|
Involvement of nuclear NHERF1 in colorectal cancer progression.
PA1-090 was used in immunohistochemistry to study the role of nuclear NHERF1 in the progression of colorectal cancer
|Mangia A,Saponaro C,Malfettone A,Bisceglie D,Bellizzi A,Asselti M,Popescu O,Reshkin SJ,Paradiso A,Simone G||Oncology reports (28:889)||2012|
Overexpression of nuclear NHERF1 in advanced colorectal cancer: association with hypoxic microenvironment and tumor invasive phenotype.
PA1-090 was used in immunohistochemistry to study the role of NHERF1 and its potential utility as a biomarker of advanced colorectal cancer
|Malfettone A,Silvestris N,Paradiso A,Mattioli E,Simone G,Mangia A||Experimental and molecular pathology (92:296)||2012|
Peritumoral vascular invasion and NHERF1 expression define an immunophenotype of grade 2 invasive breast cancer associated with poor prognosis.
PA1-090 was used in immunohistochemistry to study a category of grade 2 breast cancer defined by peritumoral vascular invasion and the expression of NHERF1
|Malfettone A,Saponaro C,Paradiso A,Simone G,Mangia A||BMC cancer (12:null)||2012|
Human epidermal growth factor receptor 2, Na+/H+ exchanger regulatory factor 1, and breast cancer susceptibility gene-1 as new biomarkers for familial breast cancers.
PA1-090 was used in immunohistochemistry to investigate the changes of gene expression in familial breast cancers
|Mangia A,Malfettone A,Saponaro C,Tommasi S,Simone G,Paradiso A||Human pathology (42:1589)||2011|
Na+/H+ exchanger regulatory factor 1 expression levels in blood and tissue predict breast tumour clinical behaviour.
PA1-090 was used in immunohistochemistry to study the prognostic value of sodium/hydrogen exchanger regulatory factor 1 for breast tumour
|Bellizzi A,Mangia A,Malfettone A,Cardone RA,Simone G,Reshkin SJ,Paradiso A||Histopathology (58:1086)||2011|
Biological role of NHERF1 protein expression in breast cancer.
PA1-090 was used in immunohistochemistry to investigate the role of sodium/hydrogen exchanger regulatory factor in tumorigenesis
|Mangia A,Chiriatti A,Bellizzi A,Malfettone A,Stea B,Zito FA,Reshkin SJ,Simone G,Paradiso A||Histopathology (55:600)||2009|
Expression and clinicopathological significance of oestrogen-responsive ezrin-radixin-moesin-binding phosphoprotein 50 in breast cancer.
PA1-090 was used in immunohistochemistry to study the expression of ezrin-radixin-moesin-binding phosphoprotein 50 in breast cancer and its clinical relevance
|Song J,Bai J,Yang W,Gabrielson EW,Chan DW,Zhang Z||Histopathology (51:40)||2007|
Expression of ion transport-associated proteins in human efferent and epididymal ducts.
PA1-090 was used in immunohistochemistry to investigate the expression and localization of ion transport-associated proteins in epididymal and efferent ducts
|Kujala M,Hihnala S,Tienari J,Kaunisto K,Hästbacka J,Holmberg C,Kere J,Höglund P||Reproduction (Cambridge, England) (133:775)||2007|
The tight-junction protein claudin-6 induces epithelial differentiation from mouse F9 and embryonic stem cells.
PA1-090 was used in immunocytochemistry and western blot to study the role of Cldn6 in epithelial differentiation.
|Sugimoto K,Ichikawa-Tomikawa N,Satohisa S,Akashi Y,Kanai R,Saito T,Sawada N,Chiba H||PloS one (8:null)||2013|
Na+/H+ exchanger regulatory factor 1 (NHERF1) directly regulates osteogenesis.
PA1-090 was used in immunocytochemistry to study the expression of NHERF1 by mineralizing osteoblasts and its role in osteogenesis
|Liu L,Alonso V,Guo L,Tourkova I,Henderson SE,Almarza AJ,Friedman PA,Blair HC||The Journal of biological chemistry (287:43312)||2012|
Receptor activity-modifying protein (RAMP) isoform-specific regulation of adrenomedullin receptor trafficking by NHERF-1.
PA1-090 was used in immunocytochemistry to demonstrate the role of NHERF-1 and RAMP3 on receptor internalization.
|Bomberger JM,Spielman WS,Hall CS,Weinman EJ,Parameswaran N||The Journal of biological chemistry (280:23926)||2005|
Foxj1 is required for apical localization of ezrin in airway epithelial cells.
PA1-090 was used in immunocytochemistry to investigate the role of Foxj1 in the apical localization of ezrin in airway epithelial cells.
|Huang T,You Y,Spoor MS,Richer EJ,Kudva VV,Paige RC,Seiler MP,Liebler JM,Zabner J,Plopper CG,Brody SL||Journal of cell science (116:4935)||2003|
PTEN, NHERF1 and PHLPP form a tumor suppressor network that is disabled in glioblastoma.
PA1-090 was used in western blot to study the regulation of phosphatidylinositol-3-OH kinase-Akt pathway in glioblastoma
|Molina JR,Agarwal NK,Morales FC,Hayashi Y,Aldape KD,Cote G,Georgescu MM||Oncogene (31:1264)||2012|
A hierarchy of signals regulates entry of membrane proteins into the ciliary membrane domain in epithelial cells.
PA1-090 was used in western blot to investigate the mechanism for the exclusion of podocalyxin from the ciliary membrane domain
|Francis SS,Sfakianos J,Lo B,Mellman I||The Journal of cell biology (193:219)||2011|
Quantitative protein and mRNA profiling shows selective post-transcriptional control of protein expression by vasopressin in kidney cells.
PA1-090 was used in western blot to study the post-transcriptional control of protein expression by vasopressin in kidney cells
|Khositseth S,Pisitkun T,Slentz DH,Wang G,Hoffert JD,Knepper MA,Yu MJ||Molecular & cellular proteomics : MCP (10:null)||2011|
Na/H exchanger regulatory factors control parathyroid hormone receptor signaling by facilitating differential activation of G(alpha) protein subunits.
PA1-090 was used in western blot to investigate the mechanism of parathyroid hormone receptor pathway modulation mediated by Na/H exchanger regulatory factors
|Wang B,Ardura JA,Romero G,Yang Y,Hall RA,Friedman PA||The Journal of biological chemistry (285:26976)||2010|
Hepatocyte nuclear factor 4alpha, a key factor for homeostasis, cell architecture, and barrier function of the adult intestinal epithelium.
PA1-090 was used in western blot to study the role of hepatocyte nuclear factor 4 in the adult mouse small intestine.
|Cattin AL,Le Beyec J,Barreau F,Saint-Just S,Houllier A,Gonzalez FJ,Robine S,Pinçon-Raymond M,Cardot P,Lacasa M,Ribeiro A||Molecular and cellular biology (29:6294)||2009|
Proteomic analysis of peripheral blood mononuclear cells: selective protein processing observed in patients with rheumatoid arthritis.
PA1-090 was used in western blot to investigate the changes of proteins in peripheral blood mononuclear cells from patients with rheumatoid arthritis
|Schulz M,Dotzlaw H,Mikkat S,Eggert M,Neeck G||Journal of proteome research (6:3752)||2007|
Activation-independent parathyroid hormone receptor internalization is regulated by NHERF1 (EBP50).
PA1-090 was used in western blot to study the effect of NHERF1 on the conditional efficacy of PTH fragments.
|Sneddon WB,Syme CA,Bisello A,Magyar CE,Rochdi MD,Parent JL,Weinman EJ,Abou-Samra AB,Friedman PA||The Journal of biological chemistry (278:43787)||2003|
EBP50; Ezrin-Radixin-Moesin Binding Phosphoprotein 50; ezrin-radixin-moesin binding phosphoprotein-50; ezrin-radixin-moesin-binding phosphoprotein 50; Na(+)/H(+) exchange regulatory cofactor NHE-RF; Na(+)/H(+) exchange regulatory cofactor NHE-RF1; Na+/H+ exchange regulatory co-factor; NHERF; regulatory cofactor of Na(+)/H(+) exchanger; So; sodium-hydrogen exchanger regulatory factor 1; solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulator 1; solute carrier family 9 (sodium/hydrogen exchanger), isoform 3 regulatory factor 1; solute carrier family 9 isoform A3 regulatory factor 1; solute carrier family 9, subfamily A (NHE3, cation proton antiporter 3), member 3 regulator 1
EBP-50; EBP50; NHE-RF; NHERF; NHERF-1; NHERF1; NPHLOP2; SLC9A3R1