Invitrogen

NEDD4 Polyclonal Antibody

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This Antibody was verified by KnockDown to ensure that the antibody binds to the antigen stated. View Details

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NEDD4 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of NEDD4 was performed using 70% confluent log phase NIH/3T3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with NEDD4 Rabbit Polyclonal Antibody(Product # PA5-17463) at 1:50 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

NEDD4 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of NEDD4 in untreated C6 cells using a NEDD4 polyclonal antibody (Product # PA5-17463) (green). DNA is labeled using a fluorescent blue dye.

NEDD4 Antibody in Western Blot (WB) — Knockdown of NEDD4 was achieved by transfecting Caco-2 with NEDD4 specific siRNAs (Silencer® select Product # s9416). Western blot analysis (Fig. a) was performed using whole cell extracts from the NEDD4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with NEDD4 Polyclonal Antibody (Product # PA5-17463, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to NEDD4.

NEDD4 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of NIH/3T3 (Lane 1), A549 (Lane 2), K-562 (Lane 3), PANC-1 (Lane 4), tissue extracts (30 µg lysate) Mouse Skeletal Muscle (Lane 5) and Rat Skeletal Muscle (Lane 6). The blot was probed with Anti-NEDD4 Polyclonal Antibody (Product # PA5-17463, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 110 kDa band corresponding to NEDD4 was observed across the cell lines and tissue extracts tested.

NEDD4 Antibody in Western Blot (WB) — Western blot analysis of NEDD4 in lysates from NIH/3T3 and C6 cells using NEDD4 polyclonal antibody (Product # PA5-17463).

NEDD4 Antibody in KnockDown — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. A549 cells were transfected with NEDD4 siRNA and reduction of signal was observed in Western Blot using NEDD4 Polyclonal Antibody (Product # PA5-17463). {KD}

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

1:100

Immunocytochemistry (ICC)

1:50

Immunofluorescence (IF)

1:50

Western Blot (WB)

1:2000

Product Specifications

Species Reactivity

Human, Mouse, Non-human primate, Rat

Host / Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Synthetic peptide corresponding to human NEDD4 protein

Conjugate

Unconjugated

Form

Liquid

Purification

Antigen affinity chromatography

Storage buffer

0.01M HEPES, pH 7.5, with 0.15M NaCl, 100µg/mL BSA, 50% glycerol

Contains

no preservative

Storage conditions

-20°C

RRID

AB_10976819

Product Specific Information

It is not recommended to aliquot this antibody.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Cell proliferation-inducing gene 53 protein; E3 ubiquitin-protein ligase NEDD4; HECT-type E3 ubiquitin transferase NEDD4; NEDD-4; Neural precursor cell expressed developmentally down-regulated protein 4; neural precursor cell expressed, developmentally down-regulated gene 4; neural precursor cell expressed, developmentally down-regulated gene 4A; neural precursor cell expressed, developmentally down-regulted gene 4; OTTHUMP00000217840; OTTHUMP00000217841; OTTHUMP00000217842; receptor-potentiating factor 1

Gene Aliases: AA959633; AL023035; AU019897; E430025J12Rik; KIAA0093; mKIAA0093; Nedd-4; NEDD4; NEDD4-1; Nedd4a; PIG53; RPF1

UniProt ID: (Human) P46934, (Rat) Q62940, (Mouse) P46935

Entrez Gene ID: (Human) 4734, (Rat) 25489, (Mouse) 17999

Molecular Function: ligase ubiquitin-protein ligase

adaptive immune response outflow tract morphogenesis endocardial cushion development protein monoubiquitination protein targeting to lysosome lysosomal transport neuromuscular junction development negative regulation of sodium ion transport negative regulation of transcription from RNA polymerase II promoter in response to UV-induced DNA damage positive regulation of phosphatidylinositol 3-kinase signaling response to denervation involved in regulation of muscle adaptation regulation of macroautophagy protein ubiquitination transmission of virus negative regulation of vascular endothelial growth factor receptor signaling pathway neuron projection development receptor internalization receptor catabolic process cellular response to UV regulation of ion transmembrane transport T cell activation regulation of membrane potential protein ubiquitination involved in ubiquitin-dependent protein catabolic process glucocorticoid receptor signaling pathway ubiquitin-dependent protein catabolic process via the multivesicular body sorting pathway development involved in symbiotic interaction positive regulation of protein catabolic process positive regulation of nucleocytoplasmic transport blood vessel morphogenesis regulation of dendrite morphogenesis regulation of synapse organization progesterone receptor signaling pathway response to calcium ion protein K63-linked ubiquitination regulation of potassium ion transmembrane transporter activity negative regulation of sodium ion transmembrane transporter activity negative regulation of transcription from RNA polymerase II promoter immune response nervous system development

Process(es)

protein binding protein C-terminus binding ligase activity sodium channel inhibitor activity protein domain specific binding beta-2 adrenergic receptor binding ubiquitin binding phosphoserine binding phosphothreonine binding ubiquitin protein ligase activity RNA polymerase binding proline-rich region binding ubiquitin-protein transferase activity UDP-N-acetylmuramoylalanyl-D-glutamyl-2,6-diaminopimelate-D-alanyl-D-alanine ligase activity ribosomal S6-glutamic acid ligase activity coenzyme F420-0 gamma-glutamyl ligase activity coenzyme F420-2 alpha-glutamyl ligase activity protein-glycine ligase activity protein-glycine ligase activity, initiating protein-glycine ligase activity, elongating tubulin-glycine ligase activity protein-glutamic acid ligase activity tubulin-glutamic acid ligase activity