General description
Na(+)/H(+) exchange regulatory cofactor NHE-RF1 (UniProt: O14745; also known as NHERF-1, Ezrin-radixin-moesin-binding phosphoprotein 50, EBP50, Regulatory cofactor of Na(+)/H(+) exchanger, Sodium-hydrogen exchanger regulatory factor 1, Solute carrier family 9 isoform A3 regulatory factor 1) is encoded by the SLC9A3R1 (also known as NHERF, NHERF1) gene (Gene ID: 9368) in human. EBP50 is a 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. Its expression is observed in several tissues, including liver, kidney, pancreas, prostate, spleen, placenta, and small intestine. It contains two PDZ domains (aa 14-94 and 154-234) that are essential for its binding to multiple ligands. It is involved in the regulation of phosphate reabsorption in the renal proximal tubules. It also plays a role in the regulation of the activity and subcellular location of Na(+)/H(+) exchanger 3 (SLC9A3) and is essential for cAMP-mediated phosphorylation and inhibition of SLC9A3. It is also reported to be essential for recycling of internalized adrenoceptor β2.Mutations in SLC9A3R1 gene are known to cause hypophosphatemic nephrolithiasis, a disease characterized by decreased renal phosphate absorption, renal phosphate wasting, hypercalciuria, and osteoporosis. Two isoforms of EBP50 have been described that are produced by alternative splicing. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Courbebaisse, M., et al. (2012). PLoS One. 7(4); e34764; Garbett, D., and Bretscher, A. (2012). J. Cell Biol. 198(2); 195-203; Cao, TT., et al. (1999). Nature. 401(6750); 286-290).
Immunogen
KLH-conjugated linear peptide corresponding to 20 amino acids from the N-terminal half of human EBP50/NHERF-1.
Application
Quality Control Testing
Evaluated by Western Blotting in Human kidney tissue lysate.
Western Blotting Analysis: A 1:10,000 dilution of this antibody detected EBP/NHERF-1 in Human kidney tissue lysate.
Tested Applications
Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected EBP/NHERF-1 in lysates from HEK293 cells, Mouse kidney tissue and Rat kidney tissue and 1:10,000 dilution detected EBP/NHERF-1 in MCF-7 cell lysate.
Affinity Binding Assay: A representative lot of this antibody bound EBP/NHERF-1 peptide with a KD of 4.9 x 10-8 in an affinity binding assay.
Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected EBP/NHERF-1 in MCF-7 cells.
Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected EBP/NHERF-1 in human kidney tissue sections.
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone 1G10 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects EBP50/NHEF-1. It targets an epitope within 20 amino acids from the N-terminal half.
Physical form
Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.
Preparation Note
Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.
Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.
Legal Information
ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
| biological source | rabbit |
| Quality Segment | 200 |
| recombinant | expressed in HEK 293 cells |
| conjugate | unconjugated |
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| clone | 1G10, recombinant monoclonal |
| description | 1G10 Clone |
| product line | ZooMAb® learn more |
| form | lyophilized |
| mol wt | calculated mol wt ~35 kDa |
| purified by | using Protein A |
| species reactivity | mouse, human, rat |
| species reactivity (predicted by homology) | monkey |
| packaging | antibody small pack of 25 μL |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry. |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation |
| sustainability | Greener Alternative Product |
| technique(s) | affinity binding assay: suitable,immunocytochemistry: suitable,immunohistochemistry: suitable,western blot: suitable |
| isotype | IgG |
| epitope sequence | N-terminal half |
| Protein ID accession no. | NP_004243 |
| UniProt accession no. | O14745 |
| greener alternative category | , Aligned |
| shipped in | ambient |
| storage temp. | 2-8°C |
| target post-translational modification | unmodified |
| Gene Information | human ... SLC9A3R1(9368) |
