New England Biolabs Canada
 

Product Pathways - Protein Stability

Skp1 Antibody #2156

Item# Description List Price Web Price Qty
2156S Skp1 Antibody - 100 µl $372.00
$334.80
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2156T Skp1 Antibody - 20 µl $159.00
$143.10
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Source
W Human, Mouse, Rat Endogenous 19 Rabbit
IF-IC
F

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry

Protocols

Specificity / Sensitivity

Skp1 Antibody detects endogenous levels of total Skp1 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues of human Skp1. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 and NIH/3T3 cells, using Skp1 Antibody.

IF-IC

IF-IC

Confocal immunofluorescent images of NIH/3T3 cells labeled with Skp1 Antibody (green, left) compared to an isotype control (right). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of NIH/3T3 cells using Skp1 antibody (blue) compared to a nonspecific negative control antibody (red).


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

  • Yamada, K. et al. (2011) J Biol Chem 286, 44057-66. Applications: Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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