Recombinant Anti-Cdk4 antibody [EPR2513Y] (ab68266)
![Western blot - Anti-Cdk4 antibody [EPR2513Y] (ab68266)](https://www.abcam.com/ps/products/68/ab68266/Images/ab68266-471157-AntiCdk4antibodyEPR2513Ywesternblothelak562ramoshumanlungratmouse.jpg "Western blot - Anti-Cdk4 antibody [EPR2513Y] (ab68266)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2513Y] to Cdk4
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cdk4 antibody [EPR2513Y]
See all Cdk4 primary antibodies -
Description
Rabbit monoclonal [EPR2513Y] to Cdk4 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide corresponding to Human Cdk4 aa 1-100 (N terminal).
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Positive control
- WB: HeLa, Ramos, and K562 cell lysates. Rat and mouse lung tissue lysates. ICC/IF: MCF7 cells. Flow Cyt (intra) (Intra): A549 cells. IP: K562 whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2513Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68266 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt (Intra) |
1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC/IF |
1/100.
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| WB |
1/500 - 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
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| IP |
1/30.
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| Notes |
|---|
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Flow Cyt (Intra)
1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/100. |
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WB
1/500 - 1/1000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). |
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IP
1/30. |
Target
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Function
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. -
Involvement in disease
Defects 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 similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. -
Post-translational
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 localization
Cytoplasm. 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
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Database links
- Entrez Gene: 1019 Human
- Entrez Gene: 12567 Mouse
- Entrez Gene: 94201 Rat
- Omim: 123829 Human
- SwissProt: P11802 Human
- SwissProt: P30285 Mouse
- SwissProt: P35426 Rat
- Unigene: 95577 Human
see all -
Alternative names
- Cdk 4 antibody
- cdk4 antibody
- CDK4 protein antibody
see all
Images
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Western blot - Anti-Cdk4 antibody [EPR2513Y] (ab68266)All lanes : Anti-Cdk4 antibody [EPR2513Y] (ab68266) at 1/2000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 4 : Rat lung lysate
Lane 5 : Mouse lung lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 34 kDa
Additional bands at: 34 kDa. We are unsure as to the identity of these extra bands.Exposure times: Lane 1-3: 30 secs; Lane 4-5: 180 secs.
Blocking/Dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] (ab68266)](https://www.abcam.com/ps/products/68/ab68266/Images/ab68266-308097-anti-cdk4-antibody-epr2513y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] (ab68266)Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cdk4 with ab68266 at 1:100 (5.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% triton X-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain.
PBS instead of the primary antibody was used as the secondary antibody only control.
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![Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] (ab68266)](https://www.abcam.com/ps/products/68/ab68266/Images/ab68266-308107-anti-cdk4-antibody-epr2513y-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] (ab68266)ab68266 at 1:30 dilution (2 µg) immunoprecipitating Cdk4 in K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate.
Lane 1 (input): K562 whole cell lysate 10 µg
Lane 2 (+): ab68266 & K562 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68266 in K562 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
![Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] (ab68266)](https://www.abcam.com/ps/products/68/ab68266/Images/ab68266-308100-anti-cdk4-antibody-epr2513y-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] (ab68266)Intracellular Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling Cdk4 with ab68266 at 1/50 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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![Anti-Cdk4 antibody [EPR2513Y] (ab68266)](https://www.abcam.com/ps/products/68/ab68266/Images/ab68266-8-benefits-of-recombinant-antibodies.png)
Anti-Cdk4 antibody [EPR2513Y] (ab68266)
Protocols
Datasheets and documents
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Datasheet download
References (16)
ab68266 has been referenced in 16 publications.
- Lv T et al. Knockdown of lncRNA DLEU1 inhibits the tumorigenesis of oral squamous cell carcinoma via regulation of miR-149-5p/CDK6 axis. Mol Med Rep 23:N/A (2021). PubMed: 33880596
- Zhang Y et al. LncRNA-CCDC144NL-AS1 Promotes the Development of Hepatocellular Carcinoma by Inducing WDR5 Expression via Sponging miR-940. J Hepatocell Carcinoma 8:333-348 (2021). PubMed: 33977095
- Stegmann KM et al. The folate antagonist methotrexate diminishes replication of the coronavirus SARS-CoV-2 and enhances the antiviral efficacy of remdesivir in cell culture models. Virus Res 302:198469 (2021). PubMed: 34090962
- Baptista Moreno Martin AC et al. [10]-Gingerol improves doxorubicin anticancer activity and decreases its side effects in triple negative breast cancer models. Cell Oncol (Dordr) 43:915-929 (2020). PubMed: 32761561
- Wei L et al. TMPO-AS1, a Novel E2F1-Regulated lncRNA, Contributes to the Proliferation of Lung Adenocarcinoma Cells via Modulating miR-326/SOX12 Axis. Cancer Manag Res 12:12403-12414 (2020). PubMed: 33293866
