Ubiquitin antibody

Catalog No. ABIN108010

Product Details anti-Ubiquitin Antibody

Target: Ubiquitin

Reactivity: Various Species

Host: Rabbit

Clonality: Polyclonal

Conjugate: This Ubiquitin antibody is un-conjugated

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

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.

Characteristics: 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 e-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.

Purification: purified

Sterility: Sterile filtered

Immunogen: This purified antibody was prepared from rabbit serum after repeated immunizations with ubiquitin coupled to rabbit IgG using glutaraldehyde.
Immunogen Type: NativeProtein

Isotype: IgG

Application Details

Application Notes: This purified polyclonal antibody reacts with ubiquitin by ELISA and western blot. Although not tested, this antibody is likely functional in immunohistochemistry and immunoprecipitation. For detection of free Ub by western blotting use Tris-Tricine SDS-PAGE and autoclaved nitrocellulose filters after the transfer and before blocking and addition of anti-Ub antibodies. Details on western blotting procedures are found in Mimnaugh et al., (1999 and 2002).

Comment:

Gene Name: UBB
Post Translational Modification: Phosphorylation

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 1.0 mg/mL

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative: Sodium azide

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

Storage: 4 °C/-20 °C

Storage Comment: Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Expiry Date: 12 months

References for anti-Ubiquitin antibody (ABIN108010)

Mimnaugh, Bonvini, Neckers: "The measurement of ubiquitin and ubiquitinated proteins." in: Electrophoresis, Vol. 20, Issue 2, pp. 418-28, (1999) (PubMed).

Target Details for Ubiquitin

Target: Ubiquitin

Alternative Name: Ubiquitin (Ubiquitin Products)

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 e-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.
Synonyms: FLJ25987 antibody, MGC8385 antibody, Polyubiquitin B antibody, RPS27A antibody, UBA52 antibody, UBA80 antibody, UBB antibody, UBC antibody, UBCEP1 antibody

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