Western blot analysis was performed on whole cell extracts (30 ug lysate) of K562 (Lane 1), COS-7 (Lane 2), HEK-293 (Lane 3), HeLa (Lane 4), and PC-3 (Lane 5). The blots were probed with Anti-Cdk4 Mouse Monoclonal Antibody (Product# MA512984, 1-2 ug/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 626520, 1:4000 dilution). A 33 kDa corresponding to Cdk4 was observed across cell lines tested. The 17 kDa isoform was observed across cell line except COS-7. Additionally 60 kDa band was seen in COS-7, HeLa and PC-3. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Pig, Rat|
|Published species reactivity||Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|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.
MA5-12984 targets Cdk4 in IF, IP, and WB applications and shows reactivity with Human, mouse, Porcine, and Rat samples.
The MA5-12984 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.
IP-MS enrichment of CDK4 (LFQ intensity): CDK4 was enriched 179-fold from MCF7 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and CDK4 antibody (Part No. MA5-12984). See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Anti-tumor activity of the TRPM8 inhibitor BCTC in prostate cancer DU145 cells.
MA5-12984 was used in western blot to study prostate cancer DU145 cells and anti-tumor activity of the TRPM8 inhibitor BCTC
|Liu T,Fang Z,Wang G,Shi M,Wang X,Jiang K,Yang Z,Cao R,Tao H,Wang X,Zhou J||Oncology letters (11:182)||2016|
Knockdown of TRPM8 suppresses cancer malignancy and enhances epirubicin-induced apoptosis in human osteosarcoma cells.
MA5-12984 was used in western blot to study the overexpression of TRPM8 in human osteosarcoma and the beneficial effects of siRNA-mediated knockdown on proliferation, metastasis and epirubicin-induced cell apoptosis
|Wang Y,Yang Z,Meng Z,Cao H,Zhu G,Liu T,Wang X||International journal of biological sciences (10:90)||2014|
Recruitment of trimeric proliferating cell nuclear antigen by G1-phase cyclin-dependent kinases following DNA damage with platinum-based antitumour agents.
MA5-12984 was used in western blot to study the role of trimeric PCNA recruited to G1-phase Cdk-p21 complexes following platinum-induced DNA damage
|He G,Kuang J,Koomen J,Kobayashi R,Khokhar AR,Siddik ZH||British journal of cancer (109:2378)||2013|
The APC/C activator Cdh1 regulates the G2/M transition during differentiation of placental trophoblast stem cells.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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|
The role of p28GANK in rat oval cells activation and proliferation.
MA5-12984 was used in western blot to investigate the importance of p28GANK in the activation and proliferation of rat oval cells
|Shan YF,Zhou WP,Fu XY,Yan HX,Yang W,Liu SQ,Cao HF,Kang B,Wu MC,Wang HY||Liver international : official journal of the International Association for the Study of the Liver (26:240)||2006|
In vitro and in vivo pharmacokinetic-pharmacodynamic relationships for the trisubstituted aminopurine cyclin-dependent kinase inhibitors olomoucine, bohemine and CYC202.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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|
Critical requirement for cell cycle inhibitors in sustaining nonproliferative states.
MA5-12984 was used in immunoprecipitation to examine the role of cell cycle inhibitors in cell growth arrest
|Pajalunga D,Mazzola A,Salzano AM,Biferi MG,De Luca G,Crescenzi M||The Journal of cell biology (176:807)||2007|
Requirement for CDK4 kinase function in breast cancer.
MA5-12984 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.
MA5-12984 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.
MA5-12984 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|
Autoimmune responses to proliferating cell nuclear antigen multiprotein complexes involved in cell proliferation are strongly associated with their structure and biologic function in patients with systemic lupus erythematosus.
MA5-12984 was used in ELISA to study the mechanism of autoimmune responses to PCNA and its associated proteins in systemic lupus erythematosus
|Kogure T,Takasaki Y,Takeuchi K,Yamada H,Nawata M,Ikeda K,Matsushita M,Matsudaira R,Kaneda K,Terasawa K,Hashimoto H||Arthritis and rheumatism (46:2946)||2002|