Human/Mouse/Rat NEDD8 Antibody

Catalog #: AF4936 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4936
AF4936-SP

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Human/Mouse/Rat NEDD8 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat NEDD8 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than
2% cross‑reactivity with recombinant human NEDD9 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human NEDD8
Leu2-Gly77
Accession # Q15843

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

2 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

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

Immunohistochemistry

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NEDD8 in Human Skeletal Muscle. NEDD8 was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using 15 µg/mL Sheep Anti-Human/Mouse/Rat NEDD8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4936) overnight at 4 °C. Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot

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Detection of Human, Mouse, and Rat NEDD8 by Western Blot. Western blot shows lysates of Recombinant Human NEDD8, HeLa human cervical epithelial carcinoma cell line, Raji human Burkitt's lymphoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and NRK rat normal kidney cell line. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4936) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NEDD8 at approximately 6 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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: NEDD8

NEDD8 (Neural precursor cell-expressed developmentally down-regulated gene 8; also Rub1) is a 6-8 kDa member of the ubiquitin family of proteins. It is expressed in striated muscle, and via its C-terminal Gly, forms covalent bonds with cullin family proteins. This conjugation generates ubiquitin ligase activity that regulates cell cycle proteins. Human pro-NEDD8 is 81 amino acids (aa) in length. It contains one ubiquitin-like domain (aa 1-65), and a 4-5 aa C-terminal propeptide that is cleaved to expose a C-terminal Gly residue that is used to generate a Gly-Lys intermolecular bond. Mature human NEDD8 shows 100% aa sequence identity to NEDD8 from both mouse and canine.

Long Name

Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 8

Entrez Gene IDs

4738 (Human); 18002 (Mouse); 25490 (Rat)

Alternate Names

FLJ43224; MGC104393; MGC125896; MGC125897; NEDD8; NEDD-8; Neddylin; Neural precursor cell expressed developmentally down-regulated protein 8; neural precursor cell expressed, developmentally down-regulated 8; Rub1; Ubiquitin-like protein Nedd8