Ubiquitin antibody

Catalog No. ABIN116959

Product Details anti-Ubiquitin Antibody

Target: Ubiquitin

Reactivity: Saccharomyces cerevisiae, Mammalian

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Ubiquitin antibody is un-conjugated

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

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

Purification: Multi-step process.

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

Application Details

Application Notes: 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.

Restrictions: For Research Use only

Handling

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

Concentration: 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.

Preservative: Sodium azide

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

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

Storage: 4 °C/-20 °C

Storage Comment: 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.

Target Details for Ubiquitin

Target: Ubiquitin

Alternative Name: Ubiquitin (Ubiquitin Products)

Synonyms: CG11624 antibody, DmUb antibody, DmUbi-p63E antibody, Dmel\\CG11624 antibody, Ub antibody, Ubi-P63e antibody, Ubi-p antibody, Ubi63E antibody, Ubi63p antibody, Ubip antibody, Ubiq antibody, Ubp antibody, ubi antibody, ubi-p63E antibody, ubi63E antibody, CG18282 antibody, CG32744 antibody, CR32744 antibody, DmUbi-p5E antibody, Dmel\\CG32744 antibody, ubiquitin antibody, si:dz180g5.5 antibody, si:busm1-180g5.5 antibody, LOC100194618 antibody, UBIQ antibody, Ubiquitin antibody, LOC100282068 antibody, Ubiquitin-63E antibody, Ubiquitin-5E antibody, transmembrane and ubiquitin-like domain containing 2 antibody, polyubiquitin antibody, Putative ubiquitin family protein antibody, ubiquitin antibody, ubiquitin B antibody, UBiQuitin antibody, Ubiquitin antibody, Ubi-p63E antibody, Ubi-p5E antibody, tmub2 antibody, LOC100194618 antibody, pco110026 antibody, LOC100135798 antibody, UBB antibody, ubq-1 antibody, LOC100190766 antibody

Background: 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

Gene ID: 7314

UniProt: P0CG47

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