Hub1 Antikörper

Antigen: Hub1

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Hefe (Saccharomyces Cerevisiae)

Applikation: ELISA, Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN100576

Produktbeschreibung

Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with recombinant yeast Hub1protein.

Format: Liquid (sterile filtered)

Beschreibung: "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 secondclass include parkin, RAD23 and DSK2, are designated ubiquitin-domain proteins (UDPs). These proteins contain domains that arerelated to ubiquitin but are otherwise unrelated to each other. Incontrast to UBLs, UDPs are not conjugated to other proteins. Hub1(""Homologous to UBiquitin"") may function as a modifier (see figure 2)but its role is unclear because it lacks the double glycine motifcharacteristic for ubiquitin and ubiquitin-like modifiers. Recently cellpolarity factors Sph1 and Hbt1 have been identified as in vivo targets ofHub1 conjugation.HUB1 has close homologs in other species including humans. The human homologs is called Ubl5."

Spezifität: 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.

Anwendungen

Applikationshinweise: Recommended Dilutions: For immunoblotting a 1:500 dilution isrecommended. A 9.7 kDa band corresponding to yeast Hub1 isdetected. Most yeast cell lysates can be used as a positive controlwithout induction or stimulation. For ELISA a 1:1,000 to 1:5,000 dilution is recommended. Researchers should determine optimaltiters for other applications. Figure 1. Immunoblot of Hub1 fusion protein. Anti-Hub1 antibody generated by immunization with recombinant yeast Hub1 was tested by immunoblot against yeast lysates expressing the Hub1-GFP fusion protein and other UBL fusion proteins. All UBLs possess limited homologyto Ubiquitin and to each other, therefore it is important to know the degree of reactivity of each antibody against each UBL.

Konzentration: 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.

Lagerung: 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.

Forschungsgebiet: Proteolyse / Ubiquitin

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Immunoblot of Hub1 fusion protein. Anti-Hub1 antibody generated by immunization with recombinant yeast Hub1 was tested by immunoblot against yeast lysates expressing the Hub1-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 Hub1 using a 1:500 dilution of antibodies-online's IgG fraction of Rabbit-anti-Hub1 (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