Ubiquitin Antikörper

Produktnummer ABIN116959

Produktdetails anti-Ubiquitin Antikörper

Target: Ubiquitin

Reaktivität: Saccharomyces cerevisiae, Säugetier

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser Ubiquitin Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Enzyme Immunoassay (EIA)

Spezifität: 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. This purified polyclonal antibody reacts with Ubiquitin by ELISA and Western blot.

Aufreinigung: Multi-step process.

Immunogen: This purified antibody was prepared from Rabbit serum after repeated immunizations with Ubiquitin coupled to Rabbit IgG using glutaraldehyde.

Anwendungsinformationen

Applikationshinweise: Suitable for ELISA (1/1,000-1/5,000) and Western blot (1/200-1/1,000). Although not tested, this antibody is likely functional in Immunohistochemistry andImmunoprecipitation. For detection of free Ub by Western blotting use Tris-Tricine SDS-PAGE and autoclavednitrocellulose filters after the transfer and before blocking and addition of anti-Ubantibodies. Details on Western blotting procedures are found in Mimnaugh et al. , (1999 and 2002).
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Rekonstitution: Restore with 0.1 mL of deionized water (or equivalent).

Konzentration: 5.0 mg/mL (by UV absorbance at 280 nm)

Buffer: 0.02 M Potassium Phosphate, 0.12 M Sodium Chloride, pH 7.2 with 0.01 % (w/v) Sodium Azide as preservative.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handhabung: Avoid repeated freezing and thawing. Dilute only prior to immediate use

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung: Store vial at 2-8 °C prior to restoration. For extended storage add glycerol to 50% and then aliquot contents and freeze at -20 °C or below. Centrifuge product if not completely clear after standing at room temperature. This antibody is stable for one month at 2-8 °C as an undiluted liquid.

Details zu Ubiquitin

Target: Ubiquitin

Andere Bezeichnung: Ubiquitin (Ubiquitin Produkte)

Hintergrund: Ubiquitin (Ub) is a small, 76-residue, protein (8.5 kDa) found both as free monomer and covalently attached to itself and other proteins in eukaryotic cells. Free Ub is a very compact and stable molecule that is easily refolded after being denatured. It is therefore recommended that for detection of free Ub on Westerns, the Tris-Tricine SDS-PAGE is used and nitrocellulose filters are autoclaved after the transfer and before blocking and addition of anti-Ub antibodies. The C-terminus of ubiquitin forms an isopeptide bond with the ε-amino group of a lysine side chain in a target protein. In this way proteins can be covalently modified by the addition of ubiquitin which may alter the target protein's function. Monoubiquitination generally targets proteins for internalization, endocytosis and lysosomal degradation, or modifies the surface charge of histones and affects chromatin compaction. Conjugation of ubiquitin (Ub) involves a three-step mechanism whereby specific enzymes (or enzyme complexes) activate and covalently link Ub to their substrates. Multi-ubiquitin chains can be built up on a single lysine of the target protein, by isopeptide bond formation usually between the carboxyl group of Gly76 of one Ub with the amino group of the side chain of Lys of the preceding ubiquitin. Although Lys48 of Ub is most frequently used for multi-ubiquitin chain formation, other lysines have also been shown to form bonds with Gly76. If a chain of multiple copies of ubiquitin is attached to a protein, this appears to target the protein for degradation by the large intacellular protease known as the 26S proteasome. The process of intracellular proteasomal proteolysis is very rapid and efficient. This makes detection of certain ubiquitinated proteins extremely difficult. Several specific inhibitors of proteasomes are commercially available and have been proven to be very useful in research applications. The detection of ubiquitinated species is further complicated by the fact that the process of ubiquitination is reversible and is accomplished by highly specific proteases present in all eukaryotic cells. It is therefore recommended that cell lysates are kept on ice and the lysis buffers contain one or more of cysteine protease inhibitors (n-ethylmaleimide, E64, etc.).Synonyms: Polyubiquitin-B, UBB

Gen-ID: 7314

UniProt: P0CG47

Pathways: Mitotic G1-G1/S Phases, Ubiquitin Proteasome Pathway