Anti-NEDD4 Antibody, clone 1D20 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1695 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in subsets of tubules of human kidney sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in subsets of tubules of human kidney sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/punctate cytoplasmic/membranous staining was observed in subsets of tubules of human kidney sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic membranous staining was observed in neurons, neuropil & glial cells of human cerebral cortex sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic membranous staining was observed in neurons, neuropil & glial cells of human cerebral cortex sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1695, Anti-Nedd4, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic membranous staining was observed in neurons, neuropil & glial cells of human cerebral cortex sections.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Rat Kidney tissue lysate was probed with Cat. No. ZRB1695, Anti-NEDD4, clone 1D20 ZooMAb® Rabbit Monoclonal (1:3,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates NEDD4 (~160 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or NEDD4 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1695, Anti-NEDD4, clone 1D20 ZooMAb® Rabbit Monoclonal. The binding of this antibody to NEDD4 peptide displayed a KD of 5.3 x 10-8 whereas binding to non-specific control peptide displayed a KD of 4.8 x 10-3.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1D20, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 149.11 kDa |
| observed mol wt ~160 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001271267  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, rat |
| species reactivity (predicted by homology) | bovine, horse, canine, monkey, porcine, feline |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P46934 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Rat Kidney tissue lysate. Western Blotting Analysis: A 1:3,000 dilution of this antibody detected NEDD4 in Rat Kidney tissue lysate. Tested applications Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected NEDD4 in human cerebral cortex and human kidney tissue sections. Affinity Binding Assay: A representative lot of this antibody bound NEDD4 with a KD of 5.3 x 10-8 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 16 amino acids from the internal region of human NEDD4. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| 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. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1D20 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects NEDD4. It targets an epitope within 16 amino acids from the internal region. |
| Storage and Stability: | 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. |
| Target description: | E3 ubiquitin-protein ligase NEDD4 (UniProt: P46934; also known as EC:2.3.2.26, Cell proliferation-inducing gene 53 protein, HECT-type E3 ubiquitin transferase NEDD4, Neural precursor cell expressed developmentally down-regulated protein 4, NEDD-4) is encoded by the NEDD4 (also known as KIAA0093, NEDD4-1, PIG53) gene (Gene ID: 4734) in human. NEDD4 is a peripheral membrane protein that is recruited to the plasma membrane by GRB10. Once complexed with GRB10 and IGF1R, it follows IGF1R internalization and remains associated with early endosomes. NEDD4 can accept ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfer the ubiquitin to targeted substrates. It is shown to specifically ubiquitinate Lysine 63 in target proteins. It also monoubiquitinates IGF1R at multiple sites, thus leading to receptor internalization and degradation in lysosomes. NEDD4 is reported to polyubiquitinate the epithelial sodium channel and any mutations that disrupt association between NEDD4 and sodium channel can lead to Liddle s syndrome, a hyperaldosteronism-like hypertension disease. NEDD4 can interact with viral proteins that contain a late-budding motif P-P-P-Y and this interaction is believed to be essential for viral particle budding of many retroviruses, like HTLV-1 Gag, and MLV Gag. NEDD4 contains four WW domains (WW 1-4; aa 610-643; 767-800; 840-873; and 892-925) and one HECT domain (aa 984-1318). The WW domains mediate its interaction with PPxY motif-containing proteins. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Maspero, E., et al. (2013). Nat. Struct. Mol. Biol. 20(6); 696-701; Rauch, S., and Martin-Serrano, J. (2011). J. Virol. 85(7); 3546-3556). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |