Cell Signaling Technology

Product Pathways - Protein Stability

Skp1 (D3J4N) Rabbit mAb #12248

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Mk (Z) Endogenous 19 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Z=Zebrafish
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Skp1 (D3J4N) Rabbit mAb recognizes endogenous levels of total Skp1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Skp1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Skp1 (D3J4N) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Skp1 isoform b (hSkp1-Myc/DDK; +), using Skp1 (D3J4N) Rabbit mAb.

IP

IP

Immunoprecipitation of Skp1 from 293T cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Skp1 (D3J4N) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Skp1 (D3J4N) Rabbit mAb.


Background

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 and then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). Combinatorial interactions of different E2 and E3 proteins result in substrate specificity (4). Recent data suggests that activated E2 associates transiently with E3, and the dissociation is a critical step for ubiquitination (5). S phase kinase-associated protein 1 (Skp1) is a critical scaffold protein of the Skp1/CUL1/F-box (SCF) E3 ubiquitin ligase protein complex. Various F-box proteins (e.g., β-TrCP, Skp2) mediate an interaction with Skp1, via their defining and conserved domain of 40 amino acids, and with substrates to be ubiquitinated (e.g., β-catenin, p27) (4).

  1. Ciechanover, A. (1998) EMBO J. 17, 7151-60.
  2. Hochstrasser, M. (2000) Nat. Cell Biol. 2, E153-7.
  3. Hochstrasser, M. (2000) Science 289, 563-4.
  4. DeSalle, L.M. and Pagano, M. (2001) FEBS Lett. 490, 179-89.
  5. Deffenbaugh, A.E. et al. (2003) Cell 114, 611-22.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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