Recombinant Anti-Cdk6 antibody [EPR4515] (ab124821)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4515] to Cdk6
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Cdk6 antibody [EPR4515]
See all Cdk6 primary antibodies -
Description
Rabbit monoclonal [EPR4515] to Cdk6 -
Host species
Rabbit -
Specificity
Based on the immunogen sequence, we do not expect the antibody to cross-react with other CDK family members. No cross-reactivity testing has been performed.
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Tested applications
Suitable for: ICC/IF, WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. within Human Cdk6 aa 300 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: Q00534 -
Positive control
- WB: Jurkat, K562, HAP1, HeLa and 293T cell lysates. IHC-P: Human tonsil tissue. ICC/IF: HeLa and wild-type HAP1 cells.
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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.
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4515 -
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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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124821 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 |
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ICC/IF |
1/500.
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WB | (1) |
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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ICC/IF
1/500. |
WB
1/50000 - 1/200000. Detects a band of approximately 37 kDa (predicted molecular weight: 37 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Probably involved in the control of the cell cycle. Interacts with D-type G1 cyclins. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. - Information by UniProt
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Database links
- Entrez Gene: 1021 Human
- Omim: 603368 Human
- SwissProt: Q00534 Human
- Unigene: 119882 Human
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Alternative names
- CDK 6 antibody
- CDK6 antibody
- CDK6_HUMAN antibody
see all
Images
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All lanes : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CDK6 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDaLanes 1- 2: Merged signal (red and green). Green - ab124821 observed at 37 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab124821 was shown to react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266059 (knockout cell lysate ab257088) was used. Wild-type HeLa and CDK6 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. ab124821 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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ab124821 staining Cdk6 in wild-type HAP1 cells (top panel) and Cdk6 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 ab124821 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] (ab124821)
ab124821 at 1/100 dilution, staining Cdk6 in Formalin fixed Paraffin-embedded Human tonsil tissue by Immunohistochemistry.
Heat mediated antigen retrieval was performed with 0.01M citrate buffer, pH 6.0.
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Lanes 1-2 : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/10000 dilution
Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate
Lanes 2 & 4 & 6 : CDK6 knockout HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 37 kDaLanes 1 and 2: Green signal from target - ab124821 observed at 37 kDa
Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
ab124821 was shown to specifically react with CDK6 when CDK6 knockout samples were used. Wild-type and CDK6 knockout samples were subjected to SDS-PAGE. ab124821 and ab8226 (loading control to beta actin) were diluted at 1/10 000 and 1/1000 respectively 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 h at room temperature before imaging. -
All lanes : Anti-Cdk6 antibody [EPR4515] (ab124821) at 1/1000 dilution
Lane 1 : Jurkat cell lysate
Lane 2 : K562 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : 293T cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat-anti-rabbit HRP at 1/2000 dilution
Predicted band size: 37 kDa
Observed band size: 37 kDa -
ICC/IF image of ab124821 at 1/100 dilution, staining Cdk6 in HeLa cells.
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Overlay histogram showing HeLa cells stained with ab124821 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab124821, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (91)
ab124821 has been referenced in 91 publications.
- Kitamura S et al. Cyclin-dependent kinase 4/6 inhibitors suppress tumor growth in extramammary Paget's disease. Cancer Sci 113:802-807 (2022). PubMed: 34866279
- Liu Y et al. Activation of miR-500a-3p/CDK6 axis suppresses aerobic glycolysis and colorectal cancer progression. J Transl Med 20:106 (2022). PubMed: 35241106
- Shi M et al. Cytokeratin 13 promotes radiotherapy sensitivity of nasopharyngeal carcinoma by downregulating the MEK/ERK pathway. IUBMB Life 74:543-553 (2022). PubMed: 35426961
- Sun R et al. CDK6 Immunophenotype Implicates Potential Therapeutic Application of CDK4/6 Inhibitors in Urothelial Carcinoma. Front Oncol 12:819003 (2022). PubMed: 35463324
- Xu Y et al. Knockdown of lncRNA ENST00000603829 Inhibits the Proliferation and Invasion of Salivary Gland Pleomorphic Adenoma through Regulating Cyclin D1. Appl Bionics Biomech 2022:8805305 (2022). PubMed: 35528529