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 (321500) 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|
|Host / Isotype||Mouse / IgG2b, kappa|
|Immunogen||This antibody was prepared against full-length, recombinant, human cyclin E protein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Gel Shift (GS)||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.
Caspases uncouple p27(Kip1) from cell cycle regulated degradation and abolish its ability to stimulate cell migration and invasion.
32-1500 was used in western blot to investigate proteolytic processing of p27
|Podmirseg SR,Jäkel H,Ranches GD,Kullmann MK,Sohm B,Villunger A,Lindner H,Hengst L||Oncogene (35:4580)||2016|
|Human||Not Cited||Evolutionary hypothesis of telomere length in primary breast cancer and brain tumour patients: a tracer for genomic-tumour heterogeneity and instability.||Mehdipour P,Kheirollahi M,Mehrazin M,Kamalian N,Atri M||Cell biology international (35:915)||2011|