Recombinant Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)
![Western blot - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-311916-anti-cdk4-antibody-epr2513y-western-blot.jpg "Western blot - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)")
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IP, ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free
See all Cdk4 primary antibodies -
Description
Rabbit monoclonal [EPR2513Y] to Cdk4 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), IP, ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: WT HAP1 whole cell lysate.
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General notes
ab236042 is the carrier-free version of ab68266.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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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 ab236042 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) |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IP |
Use at an assay dependent concentration.
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| ICC/IF |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IP
Use at an assay dependent concentration. |
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ICC/IF
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). |
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] - BSA and Azide free (ab236042)
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 1 and 2: Green signal from target - ab68266 observed at 34 kDa
Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
Unpurified ab68266 was shown to specifically react with CDK4 when CDK4 knockout samples were used. Wild-type and CDK4 knockout samples were subjected to SDS-PAGE. ab68266 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 1/10000 dilution for 1 h 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 (ab68266).
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![Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-328231-anti-cdk4-antibody-epr2513y-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)ab68266 (purified) at 1:30 dilution (2µg) immunoprecipitating Cdk4 in K562 whole cell lysate.
Lane 1 (input): K562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate 10µg
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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68266).
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![Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-328230-anti-cdk4-antibody-epr2513y-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)Intracellular Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labeling Cdk4 with purified ab68266 at 1/50 dilution (1 µg/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). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68266).
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![Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-328229-anti-cdk4-antibody-epr2513y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Cdk4 with Purified ab68266 at 1:100 (5.4 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with None. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 ?g/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68266).
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![Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-328228-anti-cdk4-antibody-epr2513y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)Unpurified ab68266 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 ab68266 at 1/250 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab68266).
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![Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)](https://www.abcam.com/ps/products/236/ab236042/Images/ab236042-6-benefits-of-recombinant-antibodies.png)
Anti-Cdk4 antibody [EPR2513Y] - BSA and Azide free (ab236042)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab236042 has not yet been referenced specifically in any publications.
