Immunofluorescence analysis of Cyclin E was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with Cyclin E Mouse Monoclonal Antibody (321600) at 2 µg - 4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 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® 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization of Cyclin E. Panel e shows no primary antibody. The images were captured at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Full-length, recombinant, human cyclin E protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
|Immunoprecipitation (IP)||1 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.
Cyclin E is a regulatory subunit of Cdk2 and controls G1 / S transition during the mammalian cell cycle. Multiple isoforms of Cyclin E are expressed only in tumors but not in the normal tissues, suggesting a post transcriptional regulation of Cyclin E. In vitro analyses indicated that these truncated variant isoforms of Cyclin E are able to phosphorylate histone H1. Alterations in Cyclin E protein have been implicated as indicators of worse prognosis in various cancers.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
||Suppression of apoptosis by PIF1 helicase in human tumor cells.||Gagou ME,Ganesh A,Thompson R,Phear G,Sanders C,Meuth M||Cancer research (71:4998)||2011|
|Human||Not Cited||Suppression of apoptosis by PIF1 helicase in human tumor cells.||Gagou ME,Ganesh A,Thompson R,Phear G,Sanders C,Meuth M||Cancer research (71:4998)||2011|
|Not Applicable||Not Cited||
A flow cytometry-based screen of nuclear envelope transmembrane proteins identifies NET4/Tmem53 as involved in stress-dependent cell cycle withdrawal.
32-1600 was used in western blot to identify nuclear envelope proteins that affect cell cycle progression
|Korfali N,Srsen V,Waterfall M,Batrakou DG,Pekovic V,Hutchison CJ,Schirmer EC||PloS one (6:null)||2011|
|Not Applicable||0.5 µg/ml||
Neisseria gonorrhoeae infection induces altered amphiregulin processing and release.
32-1600 was used in western blot to report that infection with N. gonorrhoeae causes altered amphiregulin processing and release in human epithelial cells
|Löfmark S,de Klerk N,Aro H||PloS one (6:null)||2011|
Chemical genetics approach to restoring p27Kip1 reveals novel compounds with antiproliferative activity in prostate cancer cells.
32-1600 was used in western blot to test if small molecules can restore p27 expression in cancer cells
|Rico-Bautista E,Yang CC,Lu L,Roth GP,Wolf DA||BMC biology (8:null)||2010|
|Not Applicable||1 µg/ml||
E2F4 and ribonucleotide reductase mediate S-phase arrest in colon cancer cells treated with chlorophyllin.
32-1600 was used in western blot to investigate the cellular effects of chlorophyllin treatment
|Chimploy K,Díaz GD,Li Q,Carter O,Dashwood WM,Mathews CK,Williams DE,Bailey GS,Dashwood RH||International journal of cancer (125:2086)||2009|
|Not Applicable||Not Cited||
Differential apoptosis by gallotannin in human colon cancer cells with distinct p53 status.
32-1600 was used in western blot to elucidate how treatment with gallotannin prevents cancer
|Al-Ayyoubi S,Gali-Muhtasib H||Molecular carcinogenesis (46:176)||2007|
|Not Applicable||2.6 µg/ml||
Neisseria gonorrhoeae infection causes a G1 arrest in human epithelial cells.
32-1600 was used in western blot to elucidate how Neisseria gonorrhoeae affects the cell cycle of human cells
|Jones A,Jonsson AB,Aro H||FASEB journal : official publication of the Federation of American Societies for Experimental Biology (21:345)||2007|
The F-box protein SKP2 mediates androgen control of p27 stability in LNCaP human prostate cancer cells.
32-1600 was used in western blot to assess the role of the SKP2 F-box protein in p27 regulation in prostate cancer.
|Lu L,Schulz H,Wolf DA||BMC cell biology (3:null)||2002|
|Human||Not Cited||Molecular classification and prognostication of adrenocortical tumors by transcriptome profiling.||Giordano TJ,Kuick R,Else T,Gauger PG,Vinco M,Bauersfeld J,Sanders D,Thomas DG,Doherty G,Hammer G||Clinical cancer research : an official journal of the American Association for Cancer Research (15:668)||2009|