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Immunofluorescence analysis of Cdk4 was done on 70% confluent log phase HeLa 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 Cdk4 (DCS-31 + DCS-35) Mouse Monoclonal Antibody (MA5-13720) 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 peri-nuclear 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||Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Clone||DCS-31 + DCS-35|
|Immunogen||Purified recombinant cdk4 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)||3-5 µg/10^6 cells|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||2 µg/ml|
|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.
MA5-13720 targets Cdk4 in IF, IP, and WB applications and shows reactivity with Human, mouse, and Rat samples.
The MA5-13720 immunogen is purified recombinant cdk4 protein.
A family of proteins designated as cdks are critical regulators of cell cycle progression. The prototype member of this family, p34cdc2, and a related protein cdk2, function late in the cycle while cdk4 and cdk6 are critically involved in G1 to S progression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The APC/C activator Cdh1 regulates the G2/M transition during differentiation of placental trophoblast stem cells.
MA5-13720 was used in western blot to study the role of Cdh1 in regulating G2/M transition during placental differentiation
|Naoe H,Chiyoda T,Ishizawa J,Masuda K,Saya H,Kuninaka S||Biochemical and biophysical research communications (430:757)||2013|
Regulation of keratinocyte proliferation in rats with deep, partial-thickness scald: modulation of cyclin D1-cyclin-dependent kinase 4 and histone H1 kinase activity of M-phase promoting factor.
MA5-13720 was used in western blot to study the mechanism of regulation of keratinocyte proliferation in rats with deep partial-thickness scald
|Xie T,Niu Y,Ge K,Lu S||The Journal of surgical research (147:9)||2008|
Inhibition of prostate cancer growth by muscadine grape skin extract and resveratrol through distinct mechanisms.
MA5-13720 was used in western blot to study the distinct mechanisms by which muscadine grape skin extract and resveratrol inhibit prostate cancer cell growth
|Hudson TS,Hartle DK,Hursting SD,Nunez NP,Wang TT,Young HA,Arany P,Green JE||Cancer research (67:8396)||2007|
The LxCxE pRb interaction domain of cyclin D1 is dispensable for murine development.
MA5-13720 was used in western blot to study whether the LxCxE domain of cyclin D1 which interacts with the retinoblastoma protein is required for normal murine development
|Landis MW,Brown NE,Baker GL,Shifrin A,Das M,Geng Y,Sicinski P,Hinds PW||Cancer research (67:7613)||2007|
Discovery of an oncogenic activity in p27Kip1 that causes stem cell expansion and a multiple tumor phenotype.
MA5-13720 was used in western blot to investigate the role of p27Kip1 in stem cell expansion and oncogenesis
|Besson A,Hwang HC,Cicero S,Donovan SL,Gurian-West M,Johnson D,Clurman BE,Dyer MA,Roberts JM||Genes & development (21:1731)||2007|
Cyclin D1, cdk4, and Bim are involved in thrombin-induced apoptosis in cultured cortical neurons.
MA5-13720 was used in western blot to study the mechanism of thrombin-induced apoptosis in cultured cortical neurons.
|Rao HV,Thirumangalakudi L,Desmond P,Grammas P||Journal of neurochemistry (101:498)||2007|
In vitro and in vivo pharmacokinetic-pharmacodynamic relationships for the trisubstituted aminopurine cyclin-dependent kinase inhibitors olomoucine, bohemine and CYC202.
MA5-13720 was used in western blot to study the pharmacokinetics and pharmacodynamics of three tri-substituted aminopurine Cdk inhibitors in a model of colon carcinoma
|Raynaud FI,Whittaker SR,Fischer PM,McClue S,Walton MI,Barrie SE,Garrett MD,Rogers P,Clarke SJ,Kelland LR,Valenti M,Brunton L,Eccles S,Lane DP,Workman P||Clinical cancer research : an official journal of the American Association for Cancer Research (11:4875)||2005|
p107 inhibits G1 to S phase progression by down-regulating expression of the F-box protein Skp2.
MA5-13720 was used in western blot to study the role of Skp2 in the mechanism by which p107 inhibits G1/S phase progression
|Rodier G,Makris C,Coulombe P,Scime A,Nakayama K,Nakayama KI,Meloche S||The Journal of cell biology (168:55)||2005|
Regulation and role of p21 and p27 cyclin-dependent kinase inhibitors during hepatocyte differentiation and growth.
MA5-13720 was used in western blot to study the role of the p21 and p27 cyclin-dependent kinase inhibitors in hepatocyte development and proliferation
|Ilyin GP,Glaise D,Gilot D,Baffet G,Guguen-Guillouzo C||American journal of physiology. Gastrointestinal and liver physiology (285:G115)||2003|
Interaction of Hsp90 with the nascent form of the mutant epidermal growth factor receptor EGFRvIII.
MA5-13720 was used in western blot to study the interaction of nascent EGFRvIII with Hsp90 and its importance for maintaining EGFRvIII expression
|Lavictoire SJ,Parolin DA,Klimowicz AC,Kelly JF,Lorimer IA||The Journal of biological chemistry (278:5292)||2003|
Targeted disruption of CDK4 delays cell cycle entry with enhanced p27(Kip1) activity.
MA5-13720 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|
Cyclic AMP inhibits the proliferation of thyroid carcinoma cell lines through regulation of CDK4 phosphorylation.
MA5-13720 was used in immunoprecipitation to study the role of CDK4 phosphorylation in the mechanism by which cAMP inhibits thyroid carcinoma cell proliferation
|Rocha AS,Paternot S,Coulonval K,Dumont JE,Soares P,Roger PP||Molecular biology of the cell (19:4814)||2008|
|Not Applicable||Not Cited||
Differential modification of p27Kip1 controls its cyclin D-cdk4 inhibitory activity.
MA5-13720 was used in immunoprecipitation to study if the growth state-dependent tyrosine phosphorylation of p27Kip1 determines whether or not it inhibits cdk4
|James MK,Ray A,Leznova D,Blain SW||Molecular and cellular biology (28:498)||2008|
Requirement for CDK4 kinase function in breast cancer.
MA5-13720 was used in immunoprecipitation to study the importance of cyclin D1-dependent Cdk4 kinase activity in mammary tumorigenesis
|Yu Q,Sicinska E,Geng Y,Ahnström M,Zagozdzon A,Kong Y,Gardner H,Kiyokawa H,Harris LN,Stål O,Sicinski P||Cancer cell (9:23)||2006|
p21Cip1 and p27Kip1 induce distinct cell cycle effects and differentiation programs in myeloid leukemia cells.
MA5-13720 was used in immunoprecipitation to study the role of p21Cip1 and p27Kip1 in the cell cycle and differentiation programs of myeloid leukemia cells
|Muñoz-Alonso MJ,Acosta JC,Richard C,Delgado MD,Sedivy J,León J||The Journal of biological chemistry (280:18120)||2005|
p53, p16 and cyclin D1: molecular determinants of radiotherapy treatment response in oral carcinoma.
MA5-13720 was used in immunohistochemistry to study the molecular determinants of the response to radiotherapy in oral carcinoma
|Jayasurya R,Francis G,Kannan S,Lekshminarayanan K,Nalinakumari KR,Abraham T,Abraham EK,Nair MK||International journal of cancer (109:710)||2004|
3; cell division kinase 4; cell division protein kinase 4; cyclin dependent kinase 4; cyclin-dependent kinase 4; melanoma cutaneous malignant; p34<PSK-J3>/cdk4; PSK-J3; serine/threonine kinase
CDK4; CMM3; Crk3; PSK-J3