Neural Precursor Cell Expressed, Developmentally Down-Regulated 8 (NEDD8) antibody

Antigen: Neural Precursor Cell Expressed, Developmentally Down-Regulated 8 (NEDD8)

Synonyms: Nedd-8, FLJ43224, MGC104393, MGC125896, MGC125897, Rub1, MGC105320, CG10679, DmelCG10679, nedd8, ubiquitin, si:zc14a17.5, NEDD8, GB17785, LOC100009008

Clonality: Polyclonal

Host: Rabbit

Reactivity: Yeast (Saccharomyces Cerevisiae)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN106594

Additional Information

Alternative name: Rub1 Protein (Nedd8)

Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with full-length recombinant yeast Rub1 protein.

Format: Liquid (sterile filtered)

Description: Ubiquitin-like proteins fall into two classes: the first class, ubiquitin-like modifiers (UBL's) function as modifiers in a manner analogous to that of ubiquitin. Examples of UBL's are SUMO, Rub1 (also called Nedd8), Apg12, and Hub1. Proteins of the second class include parkin, RAD23, and DSK2, and are designated ubiquitin-domain proteins (UDP's). These proteins contain domains that are related to ubiquitin but are otherwise unrelated to each other. In contrast to UBL's, UDP's are not proteolytically processed or conjugated to other proteins. Rub1, and the corresponding human homolog Nedd8, are activated by the E1 ubiquitin-activating enzyme UBA2, which forms isopeptide linkages between thioesters. Nedd8 shows 80% homology to ubiquitin. The best known targets of Rub1 modification are members of the cullin family. Cullins are subunits of an E3-ubiquitin ligase complex called the Skp1/Cul1/Cdc53-F-box (SCF). The SCF promotes transfer of ubiquitin from a ubiquitin conjugating enzyme (E2) to the target protein. Rub1 modification may regulate SCF function or localization.

Specificity: This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Application Details

Application Notes: Recommended Dilutions: For immunoblotting, a 1:500 dilution is recommended. A 6 kDa band corresponding to yeast Rub1 is detected. Most yeast cell lysates can be used as a positive control without induction or stimulation. For ELISA, a 1:1,000 to 1:5,000 dilution is recommended. Researchers should determine optimal titers for other applications. Note: This purified polyclonal antibody reacts with yeast Rub1 by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation.

Concentration: 1.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to restoration. Restore with 0.5 ml of deionized water (or equivalent). For extended storage, aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Restrictions: For Research Use only

Images

Immunoblot of Rub1 fusion protein. Anti-Rub1 antibody, generated by immunization with full-length recombinant yeast Rub1, was tested by immunoblot against a yeast cell lysate. A dilution of the antibody between 1:200 and 1:1,000 will show strong reactivity specifically with free Rub1 protein (indicated by arrow) and Rub1 conjugates. In this blot the antibody was used at a 1:500 dilution incubated overnight at 4