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Immunofluorescence analysis of SKP2 was performed using 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SKP2 Rabbit Polyclonal Antibody (511900) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant, full-length human p45SKP2 protein.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||5 ug|
|Western Blot (WB)||1-2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
Skp2 (S-phase Kinase-associated Protein 2) belongs to the family of F-box proteins that interact with the Cyclin A-Cdk2 complex. Skp2 is essential for the G1-S transition in both transformed cells and diploid fibroblasts. Biochemical and genetic experiments have demonstrated that Skp2 is required for the ubiquitination and consequent degradation of p27 in cultured mammalian cells and in vitro reconstitution assays. In normal tissues, Skp2 is expressed in tonsil and placenta. In tumor tissues, Skp2 is expressed widely, in some colon, prostate, pancreas, and skin cancers, with high expression in lung, breast, ovarian, and endometrial cancers. Research suggests that Skp2 plays a role in oncogenesis, particularly in the pathogenesis of lymphomas and breast cancers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Skp2 regulates Myc protein stability and activity.
51-1900 was used in western blot to report that Skp2 regulates Myc ubiquitination and stability.
|Kim SY,Herbst A,Tworkowski KA,Salghetti SE,Tansey WP||Molecular cell (11:1177)||2003|
|Inhibition of F-Box protein p45(SKP2) expression and stabilization of cyclin-dependent kinase inhibitor p27(KIP1) in vitamin D analog-treated cancer cells.||Lin R,Wang TT,Miller WH,White JH||Endocrinology (144:749)||2003|
|Human||Not Cited||The pRb-related protein p130 is regulated by phosphorylation-dependent proteolysis via the protein-ubiquitin ligase SCF(Skp2).||Tedesco D,Lukas J,Reed SI||Genes & development (16:2946)||2002|
CDK2/cyclin A-associated protein p45; F-box/LRR-repeat protein 1; FBXL1; p45skp2; S-phase kinase-associated protein 2 (p45); SKP2
FBL1; FBXL1; FLB1; p45; SKP2