Branched Ubiquitin Antibody Sampler Kit #33959
Product Information
Kit Usage Information
Protocols
- 3936: Western Blotting, Immunohistochemistry (Paraffin)
- 5621: Western Blotting
- 7074: Western Blotting
- 7076: Western Blotting
- 8081: Western Blotting
Product Description
Branched Ubiquitin Antibody Sampler Kit provides an economical means of detecting total and common branch specific forms of ubiquitin. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
Specificity / Sensitivity
Ubiquitin (P4D1) Mouse mAb detects ubiquitin, polyubiquitin and ubiquitinated proteins. This antibody may cross-react with recombinant NEDD8. Linkage specific antibodies K48-linkage Specific Polyubiquitin (D9D5C6) Rabbit mAb and K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb detects polyubiquitin chains formed by Lys48 and Lys63 residue linkages, respectively. These antibodies do not react with monoubiquitin or polyubiquitin chains formed by specific linkage to different lysine residues.
Source / Purification
Ubiquitin (P4D1) Mouse mAb is produced by immunizing animals with a recombinant protein expression amino acids 1-76 of full length bovine ubiquitin. Branch specific monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residus surrounding the Lys48 and Lys63 branches of the human diubiquitin chain.
Background
Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25A, and Bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (4-7).
Substrate proteins are linked to ubiquitin using seven distinct ubiquitin lysine residues (Lys6, Lys11, Lys27, Lys29, Lys33, Lys48 and Lys63). Formation of a polyubiquitin chain occurs when a lysine residue of ubiquitin is linked to the carboxy-terminal glycine of another ubiquitin. Proteins polyubiquinated at specific lysine residues display a tendency to be targeted for different processes; K48-linked polyubiquitin chains mainly target proteins for proteasomal degradation while K63-linked polyubiquitin regulates protein function, subcellular localization, or protein-protein interactions (8). K63-linked polyubiquitin chains exert nonproteolytic functions in vivo, such as protein trafficking, kinase/phosphatase activation, and DNA damage control, all of which might be important in regulation of cancer survival and development (9,10).
Substrate proteins are linked to ubiquitin using seven distinct ubiquitin lysine residues (Lys6, Lys11, Lys27, Lys29, Lys33, Lys48 and Lys63). Formation of a polyubiquitin chain occurs when a lysine residue of ubiquitin is linked to the carboxy-terminal glycine of another ubiquitin. Proteins polyubiquinated at specific lysine residues display a tendency to be targeted for different processes; K48-linked polyubiquitin chains mainly target proteins for proteasomal degradation while K63-linked polyubiquitin regulates protein function, subcellular localization, or protein-protein interactions (8). K63-linked polyubiquitin chains exert nonproteolytic functions in vivo, such as protein trafficking, kinase/phosphatase activation, and DNA damage control, all of which might be important in regulation of cancer survival and development (9,10).
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