Human/Mouse/Rat NEDD8 Antibody

Name: Human/Mouse/Rat NEDD8 Antibody
Brand: R&D Systems
SKU: MAB49361
Rating: 5 (1 reviews)

Catalog #: MAB49361 1 Review Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB49361-100
MAB49361-SP

Detection of Human, Mouse, and Rat NEDD8 by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat NEDD8 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human NEDD8 in direct ELISAs and Western blots.

Source

Monoclonal Rat IgG_2B Clone # 866916

Purification

Protein A or G purified from hybridoma culture supernatant

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

Immunocytochemistry

8-25 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

Western Blot

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Detection of Human, Mouse, and Rat NEDD8 by Western Blot. Western blot shows lysates of 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 Rat Anti-Human/Mouse/Rat NEDD8 Monoclonal Antibody (Catalog # MAB49361) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). 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.

Immunocytochemistry

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NEDD8 in HeLa Human Cell Line. NEDD8 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Mouse Anti-Human/Mouse/Rat NEDD8 Polyclonal Antibody (Catalog # MAB49361) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 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: 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 the cullin family of proteins. This conjugation generates ubiquitin ligase activity that regulates cell cycle proteins. Human proNEDD8 is 81 amino acids (aa) in length and 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 identity to NEDD8 from both mouse and rat.

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