Immunofluorescent analysis of Cyclin A (green) in HeLa cells left untreated (UT, left panels) or treated with 100nM camptothecin (CPT, right panels). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Cyclin A monoclonal antibody (Product # MA1-154) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249; lower panels) for 30 minutes. Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification. Note: Red arrows indicate nuclear translocation of Cyclin A in response to treatment.
|Tested species reactivity||Bovine, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Bovine Cyclin A fusion protein|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Frozen) (IHC (F))||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Immunoprecipitation (IP)||1 µg|
|Western Blot (WB)||1:1000|
* 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|
MA1-154 detects Cyclin A in mammalian samples and has been successfully used in Immunohistochemistry (Frozen and Paraffin), Immunofluorescence, Immunoprecipitation and Western Blot procedures. Immunoprecipitation and Western Blot analysis with MA1-154 show the accumulation of a prominent band at ~50 kDa in camptothecin and hydroxyurea treated cells. In Immunofluorescence applications, MA1-154 shows nuclear translocation of Cyclin A in G2/M arrested cells as a result of camptothecin treatment.
The protein encoded by the CCNA2 gene belongs to the highly conserved cyclin family, whose members are characterized by a dramatic periodicity in protein abundance through the cell cycle. Cyclins function as regulators of CDK kinases. Different cyclins exhibit distinct expression and degradation patterns which contribute to the temporal coordination of each mitotic event. In contrast to cyclin A1, which is present only in germ cells, cyclin A2 is expressed in all tissues tested. Cyclin A binds and activates CDC2 or CDK2 kinases, and thus promotes both cell cycle G1/S and G2/M transitions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
APOLLON protein promotes early mitotic CYCLIN A degradation independent of the spindle assembly checkpoint.
MA1-154 was used in western blot to study the spindle assembly checkpoint-independent regulation of mitotic cyclin A by the APOLLON protein
|Kikuchi R,Ohata H,Ohoka N,Kawabata A,Naito M||The Journal of biological chemistry (289:3457)||2014|
|Not Applicable||Not Cited||
Differential apoptosis by gallotannin in human colon cancer cells with distinct p53 status.
MA1-154 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||Not Cited||
Disrupting Skp2-cyclin A interaction with a blocking peptide induces selective cancer cell killing.
MA1-154 was used in immunoprecipitation and western blot to report that the Skp2-cyclin A interaction has a major role in cancer cell survival
|Ji P,Sun D,Wang H,Bauzon F,Zhu L||Molecular cancer therapeutics (6:684)||2007|