Immunofluorescent analysis of Cyclin C was performed using 70% confluent log phase HeLa cells that were serum starved for 16hr. 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 Cyclin C Rabbit Polyclonal Antibody (PA5-16227) 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) 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 is untreated cell with less signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide corresponding to aa 290-303 (Cys-QGPNGSQNSSYSQS) from the C-terminal of human cyclin C|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Immunoprecipitation (IP)||10µg/mg protein lysate|
|Western Blot (WB)||2 µg/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.
PA5-16227 targets Cyclin C in immunofluorescence, immunoprecipitation, and Western blot applications and shows reactivity with Canine, Human, mouse, and Rat samples.
The PA5-16227 immunogen is a synthetic peptide corresponding to aa 290-303 (Cys-QGPNGSQNSSYSQS) from the C-terminal of human cyclin C.
Cyclin C was originally identified by a genetic screen for human and Drosophila cDNAs that complement a triple knock-out of the CLN genes in Saccharomyces cerevisiae. Cyclin C is a nuclear protein present in a multiprotein complex. It interacts both in vitro and in vivo with cdk8, a novel protein-kinase of the cdk family. It is suggested that cyclin C and cdk8 control RNA polymerase II function. Human cdk8-cyclin C might be functionally associated with the mammalian transcription apparatus, perhaps involved in relaying growth-regulatory signals.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.