Urm1 antibody

Antigen: Urm1

Clonality: Polyclonal

Host: Rabbit

Reactivity: Yeast (Saccharomyces Cerevisiae)

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

Catalog no.: ABIN108085

Additional Information

Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast Urm1protein. Related Link(s): UBL protein-protein interactions in S.cerevisiae. ( http://depts.washington.edu/sfields/yplm/data/Nature.html )

Format: Liquid (sterile filtered)

Description: Ubiquitin-like proteins fall into two classes:the first class, ubiquitin-like modifiers (UBLs) function as modifiers in amanner analogous to that of ubiquitin. Examples of UBLs are SUMO,Rub1 (also called Nedd8), Apg8 and Apg12. Proteins of the second classinclude parkin, RAD23 and DSK2, are designated ubiquitin-domainproteins (UDPs). These proteins contain domains that are related toubiquitin but are otherwise unrelated to each other. In contrast to UBLs,UDPs are not conjugated to other proteins. Urm1 is a newly identifiedubiquitin related modifier. Urm 1 is a 99-amino acid protein terminatedwith glycine-glycine. Target proteins are conjugated to Urm1 via its C-terminal glycine. Initially Urm1 forms a thioester with a novel E1-likeprotein, Uba4.

Specificity: This product is an IgGfraction antibody purified from monospecificantiserum by a multi-step process which includesdelipidation, salt fractionation and ion exchangechromatography followed by extensive dialysisagainst the buffer stated above. Assay byimmunoelectrophoresis resulted in a singleprecipitin arc against anti-Rabbit Serum.

Application Details

Application Notes: Recommended Dilutions: For immunoblotting a 1:1,000 dilution isrecommended. A 12 kDa band corresponding to yeast Urm1 isdetected. Most yeast cell lysates can be used as a positive controlwithout induction or stimulation. For ELISA a 1:2,000 to 1:10,000dilution is recommended. Researchers should determine optimaltiters for other applications. Note: This purified polyclonal antibody reacts with yeast Urm1 by western blot and ELISA. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. This antibody using the specified conditions may recognize other prominent intrinsic bands (UBLs or their conjugates). Other intrinsic bands are readily detectable in yeast lysates at lower antibody dilutions.

Concentration: 5.0 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. Restorewith 0.1 ml of deionized water (or equivalent). For extended storagealiquot contents and freeze at -20° C or below. Avoid cycles offreezing and thawing. Centrifuge product if not completely clear afterstanding at room temperature. This product is stable for severalweeks at 4° C as an undiluted liquid. Dilute only prior to immediateuse. Expiration date is one (1) year from date of restoration.

Research Area: Proteolysis / Ubiquitin

Restrictions: For Research Use only

Images

Immunoblot of Urm1 fusion protein. Anti- Urm1 antibody generated by immunization with recombinant yeast Urm1 was tested by immunoblot against yeast lysates expressing the Urm1-GFP fusion protein and other UBL fusion proteins. All UBLs possess limited homology to Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL. Panel A shows total protein staining using ponceau. Panel B shows positions of free GFP or GFP containing recombinant proteins present in each lysate preparation after reaction with a 1:1,000 dilution of antibodies-online's anti-GFP (code # ABIN100085) followed by reaction with a 1:15,000 dilution of HRP Donkey-a-Goat IgG MX (code # 605-703-125). Panel C shows specific reaction with Urm1 using a 1:1,000 dilution of antibodies-online's IgG fraction of Rabbit-anti- Urm1 (Yeast) followed by reaction with a 1:15,000 dilution of HRP Goat-a-Rabbit IgG MX (code # ABIN102010). All primary antibodies were diluted in TTBS buffer supplemented with 5% non-fat milk and incubated with the membranes overnight at 4