Immunofluorescence analysis of Cyclin D2 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 with Cyclin D2 (DCS-3.1 + DCS-5.2) Mouse Monoclonal Antibody (MA512731) 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||Avian, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a/IgG2b|
|Clone||DCS-3.1 + DCS-5.2|
|Immunogen||Purified histidine-tagged human recombinant full length cyclin D2 protein|
|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)||Assay Dependent|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||Assay Dependent|
* 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.
MA5-12731 targets Cyclin D2 in FACS, IF, and WB applications and shows reactivity with Human, mouse, and Rat samples.
The MA5-12731 immunogen is purified histidine-tagged human recombinant full length cyclin D2 protein.
Cyclin D2 is a G1 cyclin required for G1-phase progression and is a strong candidate for a proto-oncogene. cyclin D2 can phosphorylate pRB when associated with cdk4 and/or cdk6.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
De novo CCND2 mutations leading to stabilization of cyclin D2 cause megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome.
MA5-12731 was used in immunohistochemistry - paraffin section to discover how de novo mutations of CCND2 lead to cyclin D2 stabilization and megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome
|Mirzaa GM,Parry DA,Fry AE,Giamanco KA,Schwartzentruber J,Vanstone M,Logan CV,Roberts N,Johnson CA,Singh S,Kholmanskikh SS,Adams C,Hodge RD,Hevner RF,Bonthron DT,Braun KP,Faivre L,Rivière JB,St-Onge J,Gripp KW,Mancini GM,Pang K,Sweeney E,van Esch H,Verbeek N,Wieczorek D,Steinraths M,Majewski J,Boycott KM,Pilz DT,Ross ME,Dobyns WB,Sheridan EG||Nature genetics (46:510)||2014|
Smoothened controls cyclin D2 expression and regulates the generation of intermediate progenitors in the developing cortex.
MA5-12731 was used in immunohistochemistry to study the modulation of cyclin D2 expression by Smoothened and its role in the generation of neocortical intermediate progenitors
|Komada M,Iguchi T,Takeda T,Ishibashi M,Sato M||Neuroscience letters (547:87)||2013|
Selective cortical interneuron and GABA deficits in cyclin D2-null mice.
MA5-12731 was used in immunohistochemistry to study selective cortical interneuron and GABA deficits in cyclin D2-null mice
|Glickstein SB,Moore H,Slowinska B,Racchumi J,Suh M,Chuhma N,Ross ME||Development (Cambridge, England) (134:4083)||2007|
Loss of cyclin D1 impairs cerebellar development and suppresses medulloblastoma formation.
MA5-12731 was used in immunohistochemistry to study the role of cyclin D1 in cerebellar development and medulloblastoma formation
|Pogoriler J,Millen K,Utset M,Du W||Development (Cambridge, England) (133:3929)||2006|
Cyclin D2 compensates for the loss of cyclin D1 in estrogen-induced mouse uterine epithelial cell proliferation.
MA5-12731 was used in immunohistochemistry and western blot to study the ability of cyclin D2 to compensate for cyclin D1 loss in estrogen-induced mouse uterine epithelial cell proliferation
|Chen B,Pollard JW||Molecular endocrinology (Baltimore, Md.) (17:1368)||2003|
Sustained expression of the transcription factor GLIS3 is required for normal beta cell function in adults.
MA5-12731 was used in western blot to study the role of the GLIS3 transcription factor in normal pancreatic beta-cell maintenance and proliferation in response to a high fat diet in adult mice
|Yang Y,Chang BH,Chan L||EMBO molecular medicine (5:92)||2013|
GSK-3 inactivation or depletion promotes ß-cell replication via down regulation of the CDK inhibitor, p27 (Kip1).
MA5-12731 was used in western blot to study the mechanism by which GSK-3 inactivation or depletion promotes beta-cell replication
|Stein J,Milewski WM,Hara M,Steiner DF,Dey A||Islets (3:21)||2011|
Adaptive beta-cell proliferation is severely restricted with advanced age.
MA5-12731 was used in western blot to study the impact of ageing on adaptive pancreatic beta-cell proliferation
|Rankin MM,Kushner JA||Diabetes (58:1365)||2009|
Targeted disruption of CDK4 delays cell cycle entry with enhanced p27(Kip1) activity.
MA5-12731 was used in western blot to study the role of CDK4 in cell cycle regulation using targeted knockdown
|Tsutsui T,Hesabi B,Moons DS,Pandolfi PP,Hansel KS,Koff A,Kiyokawa H||Molecular and cellular biology (19:7011)||1999|
Regulation of exit from quiescence by p27 and cyclin D1-CDK4.
MA5-12731 was used in western blot to study the role of p27 and cyclin D1-CDK4 in regulating S-phase entry of quiescent NIH 3T3 cells
|Ladha MH,Lee KY,Upton TM,Reed MF,Ewen ME||Molecular and cellular biology (18:6605)||1998|
Cyclin D3 compensates for the loss of cyclin D1 during ErbB2-induced mammary tumor initiation and progression.
MA5-12731 was used in immunoprecipitation and western blot to study the ability of cyclin D3 to compensate for the loss of cyclin D1 during ErbB2-induced mammary tumor initiation and progression
|Zhang Q,Sakamoto K,Liu C,Triplett AA,Lin WC,Rui H,Wagner KU||Cancer research (71:7513)||2011|
v-Jun overrides the mitogen dependence of S-phase entry by deregulating retinoblastoma protein phosphorylation and E2F-pocket protein interactions as a consequence of enhanced cyclin E-cdk2 catalytic activity.
MA5-12731 was used in immunoprecipitation to study the mechanism by which v-Jun promotes S-phase entry in the absence of mitogens
|Clark W,Black EJ,MacLaren A,Kruse U,LaThangue N,Vogt PK,Gillespie DA||Molecular and cellular biology (20:2529)||2000|