Cell Signaling Technology

Product Pathways - Protein Folding/Stability

Ubiquitin (P4D1) Mouse mAb #3936

Applications Reactivity Source Isotype
W IHC-P IF-IC All Mouse IgG1

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: All=All species expected

Specificity / Sensitivity

Ubiquitin (P4D1) Mouse mAb detects ubiquitin, polyubiquitin and ubiquitinated proteins.

Source / Purification

Monoclonal antibody is produced by immunizing mice with 1-76 full length ubiquitin of bovine origin. Antibody is supplied in HEPES buffer with 50% glycerol and less than 0.02% sodium azide.

Western Blotting

Western Blotting

Western blot analysis of human 293 and HeLa cells untreated or treated with 50 µM MG-132 (a 26S proteosome inhibitor) for 90 minutes, using Ubiquitin (P4D1) Mouse mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical staining of ubiquitin in paraffin-embedded breast carcinoma showing nuclear and cytoplasmic localization, using Ubiquitin (P4D1) Mouse mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of untreated HeLa cells using Ubiquitin (P4D1) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


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 transfered 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 IkappaB, p53, cdc25a and Bcl-2 have been shown to be targets for the ubiquitination-proteosome process as part of regulation of cell cycle progression, differentiation, cell stress response and apoptosis (4-7).

  1. Ciechanover, A. (1998) EMBO J. 17, 7151-7160.
  2. Hochstrasser, M. (2000) Nat. Cell Biol. 2, E153-E157.
  3. Hochstrasser, M. (2000) Science 289, 563-564.
  4. Bernardi, R. et al. (2000) Oncogene 19, 2447-2454.
  5. Aberle, H. et al. (1997) EMBO J. 16, 3797-3804.
  6. Salomoni, P. and Pandolfi, P.P. (2002) Nat. Cell Biol. 4, E152-E153.
  7. Jesenberger, V. and Jentsch, S. (2002) Nat. Rev. Mol. Cell Biol. 3, 112-121.

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