Human/Mouse/Rat NEDD4 Antibody

Catalog #: AF6218 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6218
AF6218-SP

Detection of Human, Mouse, and Rat NEDD4 by Western Blot.

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Human/Mouse/Rat NEDD4 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat NEDD4 in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human NEDD4
Asn630-Gly768
Accession # P46934

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.25 µg/mL

See below

Simple Western

20 µg/mL

Jurkat human acute T cell leukemia cells, MCF‑7 human breast cancer cell line, L6 rat myoblast cells, and HT‑2 mouse T cells

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human, Mouse, and Rat NEDD4 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, HT-2 mouse T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6218) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NEDD4 at approximately 118 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Simple Western

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Detection of Human/Mouse/Rat NEDD4 by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cells, MCF‑7 human breast cancer cells, L6 rat myoblast cells, and HT‑2 mouse T cells, loaded at 0.2 mg/mL. A specific band was detected for NEDD4 at approximately 130 kDa (as indicated) using 20 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6218) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: NEDD4

NEDD4 (Neural precursor cell-expressed developmentally down-regulated gene 4-1; also RPF1) is a 115-120 kDa member of the HECT E3 ubiquitin ligase family of proteins. The NEDD4 gene is widely expressed, and serves as a complement to a related gene product termed NEDD4-2/NEDD4L. Both molecules ubiquitinate multiple proteins. NEDD4-1 acts preferentially on PTEN, tumor suppressors and endocytic proteins, while NEDD4-2 acts preferentially on membrane transporters. Human NEDD4-1 is 900 amino acids (aa) in length. It contains one PKC region (aa 20-124), four WW domains (aa 196-505) and one E3 HECT domain (aa 543‑897). Caspase cleavage after Asp197 generates a 95 kDa C-terminal, and smaller but variable sized N-terminal fragment. There are multiple splice forms. Two show a 516 aa substitution for aa 1-169 and 1-97, respectively, while a third shows the latter substitution coupled to a deletion of aa 170-185. Over aa 211-349, human NEDD4-1 shares 83% aa identity to mouse NEDD4-1 and less than 25% aa identity with human NEDD4-2.

Long Name

Neural Precursor Cell Expressed, Developmentally Down-regulated Protein 4

Entrez Gene IDs

4734 (Human); 17999 (Mouse); 25489 (Rat)

Alternate Names

Cell proliferation-inducing gene 53 protein; E3 ubiquitin-protein ligase NEDD4; EC 6.3.2; EC 6.3.2.-; KIAA0093NEDD4-1MGC176705; NEDD4; NEDD-4; Neural precursor cell expressed developmentally down-regulated protein 4; neural precursor cell expressed, developmentally down-regulated 4; receptor-potentiating factor 1; RPF1