Product nameAnti-Cdk4 antibody [EPR4513-32-7]
See all Cdk4 primary antibodies
DescriptionRabbit monoclonal [EPR4513-32-7] to Cdk4
Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Human
Synthetic peptide. within Human Cdk4 aa 250 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P11802
- WB: Wild-type HAP1 cell lysate; HeLa, MCF7, K562, and Ramos cell lysates. IHC-P: Human urothelial carcinoma and cervix carcinoma tissues. ICC/IF: Wild type HAP1 cells; MCF7 cells. Flow Cyt: MCF-7 cells.
A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Dissociation constant (KD)KD = 1.86 x 10 -11 M Learn more about KD
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
Concentration information loading...
PurityProtein A purified
- Anti-Cdk4 antibody [EPR4513-32-7] (Alexa Fluor® 488) (ab193966)
- Anti-Cdk4 antibody [EPR4513-32-7] (Alexa Fluor® 647) (ab193967)
- Anti-Cdk4 antibody [EPR4513-32-7] - Loading Control (HRP) (ab193968)
- Anti-Cdk4 antibody [EPR4513-32-7] - BSA and Azide free (ab213216)
- Anti-Cdk4 antibody [EPR4513-32-7] (Alexa Fluor® 555) (ab215318)
- HeLa nuclear extract lysate (ab14655)
- K562 nuclear extract lysate (ab14851)
- HeLa whole cell lysate (ab150035)
- MCF7 whole cell lysate (ab29537)
- MCF7 membrane extract lysate (ab29539)
- HeLa whole cell lysate (ab29545)
- HeLa membrane extract lysate (ab29547)
- MCF7 whole cell lysate (ab3871)
- Ramos whole cell lysate (ab3955)
- K562 whole cell lysate (ab7911)
Our Abpromise guarantee covers the use of ab108357 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionSer/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.
Involvement in diseaseDefects in CDK4 are a cause of susceptibility to cutaneous malignant melanoma type 3 (CMM3) [MIM:609048]. Malignant melanoma is a malignant neoplasm of melanocytes, arising de novo or from a pre-existing benign nevus, which occurs most often in the skin but also may involve other sites.
Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain.
modificationsPhosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.
Cellular localizationCytoplasm. Nucleus. Membrane. Cytoplasmic when non-complexed. Forms a cyclin D-CDK4 complex in the cytoplasm as cells progress through G(1) phase. The complex accumulates on the nuclear membrane and enters the nucleus on transition from G(1) to S phase. Also present in nucleoli and heterochromatin lumps. Colocalizes with RB1 after release into the nucleus.
- Information by UniProt
- Cdk 4 antibody
- cdk4 antibody
- CDK4 protein antibody
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 4 and 6: CDK4 knockout HAP1 cell lysate (20 µg)
Lanes 5 and 6: Merged signal (red and green). Green - ab108357 observed at 34kDa. Red - loading control to beta actin observed at 40kDa.
ab108357 was shown to specifically react with CDK4 in wild-type HAP1 cells. No band was observed when CDK4 knockout samples were examined. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab108357 and ab8226 (loading control to beta actin) were both diluted at 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling Cdk4 with purified ab108357 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer (pH 9). ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L), was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemical analysis of MCF7 (human breast adenocarcinoma cell line) cells, labeling Cdk4 with purified ab108357 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500), was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
ab108357 staining CDK4 in the human cell line MCF7 (human breast adenocarcinoma cell line) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)
All lanes : Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/10000 dilution (purified)
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : K562 (human chronic myelogenous leukemia cell line from bone marrow) cell lysate
Lane 3 : Ramos (human Burkitt's lymphoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa
Blocking and dilution buffer: 5% NFDM/TBST.
ab108357 staining Cdk4 in wild-type HAP1 cells (top panel) and Cdk4 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108357 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Flow Cytometry analysis of MCF7 (human breast adenocarcinoma cell line) cells labelling Cdk4 with purified ab108357 at 1/40 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human urothelial carcinoma tissue labelling Cdk4 with unpurified ab108357 at a dilution of 1/100.
All lanes : Anti-Cdk4 antibody [EPR4513-32-7] (ab108357) at 1/1000 dilution (unpurified)
Lane 1 : Human osteosarcoma whole cell lysate - control, non-targeting siRNA
Lane 2 : Human osteosarcoma whole cell lysate - siRNA for CDK4
Lysates/proteins at 20 µg per lane.
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 5 seconds
Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KD
This product has been referenced in:
- Fang G et al. LncRNA MRAK048635_P1 is critical for vascular smooth muscle cell function and phenotypic switching in essential hypertension. Biosci Rep 39:N/A (2019). Read more (PubMed: 30833363) »
- Shao G et al. lncRNA CASC9 positively regulates CHK1 to promote breast cancer cell proliferation and survival through sponging the miR-195/497 cluster. Int J Oncol 54:1665-1675 (2019). Read more (PubMed: 30816435) »