Rabbit Recombinant Monoclonal CDK4 antibody. Carrier free. Suitable for IP, WB, ICC/IF and reacts with Mouse, Rat, Human samples.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
IP | WB | ICC/IF | |
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Human | Expected | Tested | Tested |
Mouse | Tested | Tested | Tested |
Rat | Expected | Tested | Expected |
Species | Dilution info | Notes |
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Species Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat, Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex (By similarity).
Cyclin-dependent kinase 4, CRK3, Cell division protein kinase 4, PSK-J3, Crk3, Cdk4
Rabbit Recombinant Monoclonal CDK4 antibody. Carrier free. Suitable for IP, WB, ICC/IF and reacts with Mouse, Rat, Human samples.
Cyclin-dependent kinase 4, CRK3, Cell division protein kinase 4, PSK-J3, Crk3, Cdk4
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
Yes
EPR17525
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab236019 is the carrier-free version of Anti-Cdk4 antibody [EPR17525] ab199728.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Cdk4 also known as Cyclin-dependent kinase 4 is a serine/threonine kinase involved in cell cycle regulation. Cdk4 has a molecular weight of approximately 34 kDa. This protein mainly locates in the nucleus of the cell and its expression occurs in diverse tissues. Cdk4 partners with D-type cyclins to exert its function which is essential for the transition from the G1 phase to the S phase of the cell cycle. Researchers often study Cdk4 using techniques like immunohistochemistry (IHC) to determine its expression and localization in different tissues.
Cdk4 plays a fundamental role in cell cycle regulation by forming a complex with D-type cyclins. Together they phosphorylate the retinoblastoma protein (Rb) resulting in the release of E2F transcription factors which promote the expression of genes necessary for DNA replication. The Cdk4/cyclin D complex regulates the cell's commitment to enter S phase and continue cell division. This regulation is key for normal cell proliferation and tissue homeostasis. Cdk4 activity is strictly controlled by INK4 family members acting as inhibitors and adding an extra regulation layer.
Cdk4 is an integral part of important signaling pathways like the cell cycle and PI3K/AKT pathways. In the cell cycle pathway Cdk4 relays signals downstream that drive the transition from the G1 to S phase through its interaction with cyclin D and Rb. In the PI3K/AKT pathway signaling can influence cyclin D expression indirectly affecting Cdk4 activity. Proteins such as Cdk6 closely relate to Cdk4 and often compensate or partner with Cdk4 in these pathways to maintain proper cell cycle progression.
Abnormalities in Cdk4 function or regulation relate to cancer and certain metabolic syndromes. Hyperactivity of Cdk4 due to overexpression or mutation frequently occurs in several cancers including melanoma and breast cancer. In these malignancies Cdk4 aberrantly drives excessive cell proliferation. Mutations or alterations in proteins such as p16 an inhibitor of Cdk4 are often found in these cancers highlighting the critical role of Cdk4 in disease progression. Understanding the role of Cdk4 allows researchers to develop targeted therapies such as Cdk4/6 inhibitors offering new treatment options.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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This data was developed using the same antibody clone in a different buffer formulation (Anti-Cdk4 antibody [EPR17525] ab199728).
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk4 antibody [EPR17525] ab199728 observed at 34 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.
Anti-Cdk4 antibody [EPR17525] ab199728 was shown to react with Cdk4 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK4 knockout HeLa cell line ab255378 (knockout cell lysate Human CDK4 knockout HeLa cell lysate ab263780) was used. Wild-type HeLa and CDK4 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-Cdk4 antibody [EPR17525] ab199728 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Cdk4 antibody [EPR17525] (Anti-Cdk4 antibody [EPR17525] ab199728) at 1/2000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CDK4 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Cdk4 with Anti-Cdk4 antibody [EPR17525] ab199728 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody- Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-Cdk4 antibody [EPR17525] ab199728 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk4 antibody [EPR17525] ab199728).
Lanes 1 - 3: Merged signal (red and green). Green - Anti-Cdk4 antibody [EPR17525] ab199728 observed at 34 kDa. Red - loading control, ab18058, observed at 130 kDa.
Anti-Cdk4 antibody [EPR17525] ab199728 was shown to specifically recognize Cdk4 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Cdk4 knockout samples were examined. Wild-type and Cdk4 knockout samples were subjected to SDS-PAGE. Anti-Cdk4 antibody [EPR17525] ab199728 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk4 antibody [EPR17525] ab199728).
All lanes: Western blot - Anti-Cdk4 antibody [EPR17525] (Anti-Cdk4 antibody [EPR17525] ab199728) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: Cdk4 knockout HAP1 whole cell lysate at 20 µg
Lane 3: Wild-type HeLa whole cell lysate at 20 µg
Predicted band size: 34 kDa
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Cdk4 with Anti-Cdk4 antibody [EPR17525] ab199728 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H & L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic, nuclear and membrane staining on HeLa cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: Anti-Cdk4 antibody [EPR17525] ab199728 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk4 antibody [EPR17525] ab199728).
Cdk4 was immunoprecipitated from 1mg NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with Anti-Cdk4 antibody [EPR17525] ab199728 at 1/50 dilution. Western blot was performed from the immunoprecipitate using Anti-Cdk4 antibody [EPR17525] ab199728 at 1/1000 dilution. Anti-Rabbit IgG (HRP) specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.
Lane 1: NIH/3T3 whole cell lysate, 10 (Input).
Lane 2: Anti-Cdk4 antibody [EPR17525] ab199728 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit IgG,monoclonal [EPR25A]- Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Cdk4 antibody [EPR17525] ab199728 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk4 antibody [EPR17525] ab199728).
All lanes: Immunoprecipitation - Anti-Cdk4 antibody [EPR17525] (Anti-Cdk4 antibody [EPR17525] ab199728)
Predicted band size: 34 kDa
Observed band size: 34 kDa
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