Skp2 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

The critical role that the family of regulatory proteins known as cyclins plays in eukaryotic cell cycle regulation is well established. The best characterized cyclin complex is the mitotic cyclin B/Cdc2 p34 kinase, the active component of MPF (maturation promoting factor). Cyclin A accumulates prior to cyclin B in the cell cycle, appears to be involved in control of S phase and has been shown to associate with cyclin dependent kinase 2 (Cdk2). In addition, cyclin A has been implicated in cell transformation and is found in complexes with E1A, transcription factors DP-1 and E2F, and retinoblastoma protein p110. Two cyclin A-Cdk2 complex binding proteins, Skp1 p19 and Skp2 p45, have been described. Although the Skps (S phase kinase-associated proteins) associate with the active cyclin A-Cdk2 complex, they do not exhibit any regulatory effects on the complex. Abolition of Skp2 p45 function by either microinjection of anti-p45 antibodies or addition of antisense oligonucleotides prevents entry into S phase of both normal and transformed cells.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-15995

• Skp2 Antibodies
• Skp2 Lysate
• Skp2 Proteins