Rub1 antibody

Antigen: Rub1

Clonality: Polyclonal

Host: Rabbit

Reactivity: Yeast

Application: ELISA, Western Blotting (WB)

Catalog no.: ABIN116956

Additional Information

UniProt: Q03919

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

Cross-Reactivity: Yeast

Description: Ubiquitin-like proteins fall into two classes: the first class, ubiquitin-like modifiers (UBLs)function as modifiers in a manner analogous to that of ubiquitin. Examples of UBLs areSUMO, Rub1 (also called Nedd8), Apg12, Hub1. Proteins of the second class include parkin,RAD23 and DSK2, are designated ubiquitin-domain proteins (UDPs). These proteinscontain domains that are related to ubiquitin but are otherwise unrelated to each other. Incontrast to UBLs, UDPs are not proteolytically processed or conjugated to other proteins. Rub1 and the corresponding human homolog Nedd8 are activated by the E1ubiquitin-activating enzyme UBA2, that forms isopeptide linkages between thio esters. Nedd8 shows 80% homology to ubiquitin. The best known targets of Rub1 modification aremembers of the cullin family. Cullins are subunits of an E3-ubiquitin ligase complex calledthe Skp1-/Cul1/Cdc53-F-box (SCF). The SCF promotes transfer of ubiquitin from a ubiquitinconjugating enzyme (E2) to the target protein. Rub1 modification may regulate SCF functionor localization.

Characteristics: Synonyms: YDR139C, NEDD8-like protein RUB1, Ubiquitin-like protein RUB1

Application Details

Application Notes: Suitable for Immunoblotting (Western Blot: 1/500-1/2,000) and ELISA (1/1,000-1/5,000). Although not tested, this antibody is likely functional in Immunohistochemistry andImmunoprecipitation. 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. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.

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

Purification: Purified

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01%(w/v) Sodium Azide as preservative.Reconstitution: Restore with 0.5 ml of deionized water (or equivalent).

Storage: Store vial at 2-8°C prior to restoration. For extended storage add glycerol to 50% and thenaliquot contents and freeze at -20°C or below. Centrifuge product if not completely clearafter standing at room temperature. This antibody is stable for one month at 2-8°C as an undiluted liquid. Dilute only prior to immediate use. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

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° C in 5% non-fat dry milk in TTBS. Detection occurred using a 1:2000 dilution of HRP-labeled Donkey anti-Rabbit IgG (code # 611-703-127) for 1 hour at room temperature. A chemilumi-nescence system was used for signal detection (Roche). Other detection systems will yield similar results.

Publications

Laufs, Nigrovic, Schneider et al.: "Giant cell myocarditis in a 12-year-old girl with common variable immunodeficiency." in: Mayo Clinic proceedings. Mayo Clinic, Vol. 77, Issue 1, pp. 92-6, 2002 (PubMed).