Immunofluorescence analysis of CDC25A was done on 70% confluent log phase MCF-7 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 CDC25A (DCS-120) Mouse Monoclonal Antibody (MA513794) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified recombinant CDC25A|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||Assay Dependent|
|Immunoprecipitation (IP)||2 µg/mg protein lysate|
|Western Blot (WB)||1-2 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.
MA5-13794 targets CDC25A in IP, IHC (P) and WB applications and shows reactivity with Human and Rat samples.
The MA5-13794 immunogen is purified recombinant CDC25A.
Cyclin-dependent kinases are positively regulated by association with cyclins and negatively regulated by binding to inhibitory subunits. The activity of cyclin-dependent kinases is also regulated by the phosphorylation status, which is controlled by the antagonistic action of wee1 kinase and CDC25 phosphatases. Three CDC25 genes are present in human cells: CDC25A, CDC25B, and CDC25C. These three genes function at different phases of the cell cycle. Whereas CDC25A and CDC25B are expressed throughout the cell cycle, with peak expression in G1 for CDC25A and in both G1-S-phase and G2 for CDC25B, CDC25C is predominantly expressed in G2.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
A subset of cancer cell lines is acutely sensitive to the Chk1 inhibitor MK-8776 as monotherapy due to CDK2 activation in S phase.
MA5-13794 was used in western blot to report that inappropriate activation of CDK2 in S phase affects checkpoint kinase 1 inhibitor sensitivity in a subset of cell lines
|Sakurikar N,Thompson R,Montano R,Eastman A||Oncotarget (7:1380)||2016|
The let-7 microRNA represses cell proliferation pathways in human cells.
MA5-13794 was used in western blot to study the role of let-7 microRNA in repressing human cell proliferation pathways
|Johnson CD,Esquela-Kerscher A,Stefani G,Byrom M,Kelnar K,Ovcharenko D,Wilson M,Wang X,Shelton J,Shingara J,Chin L,Brown D,Slack FJ||Cancer research (67:7713)||2007|
Differential utilization of cyclin D1 and cyclin D3 in the distinct mitogenic stimulations by growth factors and TSH of human thyrocytes in primary culture.
MA5-13794 was used in western blot to study the roles of cyclin D1 and cyclin D3 in different mitogenic modes of human thyrocytes
|Paternot S,Dumont JE,Roger PP||Molecular endocrinology (Baltimore, Md.) (20:3279)||2006|
SCFbeta-TRCP links Chk1 signaling to degradation of the Cdc25A protein phosphatase.
MA5-13794 was used in western blot to study the role of SCFbeta-TRCP in cell cycle arrest and DNA repair
|Jin J,Shirogane T,Xu L,Nalepa G,Qin J,Elledge SJ,Harper JW||Genes and development (17:3062)||2003|
CDC25A protein stability represents a previously unrecognized target of HER2 signaling in human breast cancer: implication for a potential clinical relevance in trastuzumab treatment.
MA5-13794 was used in immunohistochemistry to study the expression of CDC25A in HER2-positive breast cancer and the correlation between CDC25A levels and trastuzumab sensitivity
|Brunetto E,Ferrara AM,Rampoldi F,Talarico A,Cin ED,Grassini G,Spagnuolo L,Sassi I,Ferro A,Cuorvo LV,Barbareschi M,Piccinin S,Maestro R,Pecciarini L,Doglioni C,Cangi MG||Neoplasia (New York, N.Y.) (15:579)||2013|