Knockout Tested Rabbit Recombinant Monoclonal CDK6 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 6 publications.
Images
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img140471.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395)")
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img120889.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395)")
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img128977.jpg/_jcr_content/renditions/webp-475.webp "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395)")
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img68864.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395)")
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img118738.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (AB222395)")
Publications
Filter by
- Evidence-based complementary and alternative medic 2016:59896812016Activation of p53/miR-34a Tumor Suppressor Axis by Chinese Herbal Formula JP-1 in A549 Lung Adenocarcinoma Cells.
- Oncology letters 12:5023-50282016Upregulated microRNA-143 inhibits cell proliferation in human nasopharyngeal carcinoma.
- Scientific reports 6:357832016Simvastatin induces cell cycle arrest and inhibits proliferation of bladder cancer cells via PPARγ signalling pathway.
- Oncology letters 12:1129-11312016Massive renal urothelial carcinoma with renal vein tumor thrombus, pancreatic infiltration and adrenal metastasis: A case report.
- Scientific reports 5:162292015ZFP36L1 promotes monocyte/macrophage differentiation by repressing CDK6.
- Molecular medicine reports 12:6491-5002015Downregulation of microRNA‑33a promotes cyclin‑dependent kinase 6, cyclin D1 and PIM1 expression and gastric cancer cell proliferation.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| IHC-P | ICC/IF | IP | WB | Flow Cyt (Intra) | |
|---|---|---|---|---|---|
| Human | Tested | Tested | Not recommended | Tested | Tested |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
IP
Not recommendedNot recommended
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Associated Products
Select an associated product type
7 products for Alternative Product
8 products for Alternative Version
Target data
Function
Serine/threonine-protein kinase involved in the control of the cell cycle and differentiation; promotes G1/S transition. Phosphorylates pRB/RB1 and NPM1. Interacts with D-type G1 cyclins during interphase at G1 to form a pRB/RB1 kinase and controls the entrance into the cell cycle. Involved in initiation and maintenance of cell cycle exit during cell differentiation; prevents cell proliferation and negatively regulates cell differentiation, but is required for the proliferation of specific cell types (e.g. erythroid and hematopoietic cells). Essential for cell proliferation within the dentate gyrus of the hippocampus and the subventricular zone of the lateral ventricles. Required during thymocyte development. Promotes the production of newborn neurons, probably by modulating G1 length. Promotes, at least in astrocytes, changes in patterns of gene expression, changes in the actin cytoskeleton including loss of stress fibers, and enhanced motility during cell differentiation. Prevents myeloid differentiation by interfering with RUNX1 and reducing its transcription transactivation activity, but promotes proliferation of normal myeloid progenitors. Delays senescence. Promotes the proliferation of beta-cells in pancreatic islets of Langerhans. May play a role in the centrosome organization during the cell cycle phases (PubMed:23918663).
Alternative names
CDKN6, CDK6, Cyclin-dependent kinase 6, Cell division protein kinase 6, Serine/threonine-protein kinase PLSTIRE
Recommended products
Knockout Tested Rabbit Recombinant Monoclonal CDK6 antibody. Carrier free. Suitable for IHC-P, ICC/IF, WB, Flow Cyt (Intra) and reacts with Human samples. Cited in 6 publications.
Key facts
- Isotype
- IgG
- Host species
- Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
- Liquid
- Clonality
- Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
- Yes
- Clone number
- EPR4515
- Purification technique
- Affinity purification Protein A
- 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.
- Concentration
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- Do Not Freeze
Notes
ab222395 is the carrier-free version of Anti-Cdk6 antibody [EPR4515] ab124821.
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.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Cdk6 also known as Cyclin-dependent kinase 6 is a protein kinase involved in the regulation of the cell cycle. It has a molecular weight of approximately 40 kDa. Cdk6 is expressed in various tissues including lymphoid tissues and the brain. It plays a mechanical role by phosphorylating target proteins which impacts cell cycle progression particularly during the transition from G1 phase to S phase. Cdk6 can work in conjunction with D-type cyclins to form a complex that is critical for its activity.
- Biological function summary
Cdk6 functions as a regulator of the cell cycle. It participates in a complex with cyclin D to control the G1 phase progression. This activity is essential for the regulation of cell division and proliferation. The association with cyclin D allows Cdk6 to phosphorylate the retinoblastoma protein (Rb) leading to the release of E2F transcription factors that promote the expression of genes necessary for DNA synthesis.
- Pathways
Cdk6 is an important component of the cell cycle control pathway. It interacts primarily with the retinoblastoma protein and Cyclin D1 in this context. The cell cycle pathway is critical for controlled cell proliferation and its dysregulation can lead to diseases like cancer. Another involved pathway is the PI3K/AKT pathway which can activate Cdk6 activity through upstream signaling events implicating Cdk6 in cellular responses to growth signals.
- Associated diseases and disorders
Cdk6 has a significant role in cancer development due to its involvement in uncontrolled cell proliferation. Its overexpression or dysregulation is often linked to cancers such as leukemia and glioblastoma. In cancer the interaction with proteins like cyclin D1 can lead to unchecked progression through the cell cycle contributing to oncogenesis. Additionally Cdk6 inhibitors are being explored as therapeutic agents in cancer treatment due to their potential to restore control over cell cycle progression.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
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.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
9 product images
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img140471.jpg "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Cdk6 antibody [EPR4515] ab124821).
Lanes 1- 2: Merged signal (red and green). Green - Anti-Cdk6 antibody [EPR4515] ab124821 observed at 37 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) observed at 50 kDa.
Anti-Cdk6 antibody [EPR4515] ab124821 was shown to react with Cdk6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human CDK6 knockout HeLa cell line ab266059 (knockout cell lysate Human CDK6 knockout HeLa 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. Anti-Cdk6 antibody [EPR4515] ab124821 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (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 (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-Cdk6 antibody [EPR4515] (Anti-Cdk6 antibody [EPR4515] ab124821) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CDK6 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDa
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img120889.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Anti-Cdk6 antibody [EPR4515] 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 Anti-Cdk6 antibody [EPR4515] ab124821 at 1/500 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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 (Anti-Cdk6 antibody [EPR4515] ab124821).
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img128977.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Anti-Cdk6 antibody [EPR4515] ab124821 at 1/100 dilution, staining Cdk6 in formalin-fixed paraffin-embedded human tonsil tissue by immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk6 antibody [EPR4515] ab124821).
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img68864.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Clone EPR4515 (ab222395) has been successfully conjugated by Abcam. This image was generated using Anti-Cdk6 antibody [EPR4515] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-Cdk6 antibody [EPR4515] ab198944 for protocol details.
Alexa Fluor® 488 Anti-Cdk6 antibody [EPR4515] ab198944 staining Cdk6 in HeLa cells. The cells were fixed with 100% methanol (5min), 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 overnight at +4°C with Alexa Fluor® 488 Anti-Cdk6 antibody [EPR4515] ab198944 at 1/100 dilution (shown in green) and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed, at 1µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img118738.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Clone EPR4515 (ab222395) has been successfully conjugated by Abcam. This image was generated using Anti-Cdk6 antibody [EPR4515] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-Cdk6 antibody [EPR4515] ab198946 for protocol details.
Alexa Fluor® 647 Anti-Cdk6 antibody [EPR4515] ab198946 staining Cdk6 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), 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 overnight at +4°C with Alexa Fluor® 647 Anti-Cdk6 antibody [EPR4515] ab198946 at 1/100 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img137149.jpg "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Lanes 1 and 2: Green signal from target - Anti-Cdk6 antibody [EPR4515] ab124821 observed at 37 kDa
Lanes 3 and 4: Red signal from loading control - Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 observed at 42 kDa
Lanes 5 and 6: Merged (red and green) signal
Anti-Cdk6 antibody [EPR4515] 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. Anti-Cdk6 antibody [EPR4515] ab124821 and Anti-beta Actin antibody [mAbcam 8226] - Loading Control 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 (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 1/10,000 dilution for 1 h at room temperature before imaging.Lanes 1 - 2: Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395) at 1/10000 dilution
Lanes 3 - 4: Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395) at 1/1000 dilution
Lanes 5 - 6: Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Lanes 1, 3 and 5: Wild-type HAP1 cell lysate at 20 µg
Lanes 2, 4 and 6: CDK6 knockout HAP1 cell lysate at 20 µg
Predicted band size: 37 kDa
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img69963.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
ICC/IF image of Anti-Cdk6 antibody [EPR4515] ab124821 at 1/100 dilution, staining Cdk6 in HeLa cells.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk6 antibody [EPR4515] ab124821).
![Flow Cytometry (Intracellular) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--flow-cytometry-intracellular-img136862.jpg "Flow Cytometry (Intracellular) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Flow Cytometry (Intracellular) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
Overlay histogram showing HeLa cells stained with Anti-Cdk6 antibody [EPR4515] 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 (Anti-Cdk6 antibody [EPR4515] ab124821, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdk6 antibody [EPR4515] ab124821).
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img433949.jpg "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Cdk6 antibody [EPR4515] ab124821).
Western blot: Anti-CDK6 antibody [EPR4515] (Anti-Cdk6 antibody [EPR4515] ab124821) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Cdk6 antibody [EPR4515] ab124821 was shown to bind specifically to CDK6. A band was observed at 37 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CDK6 knockout cell line. To generate this image, wild-type and CDK6 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Cdk6 antibody [EPR4515] (Anti-Cdk6 antibody [EPR4515] ab124821) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: CDK6 knockout HCT 116 cell lysate at 20 µg
Lane 3: Wild-type HeLa ab255929 cell lysate at 20 µg
Lane 4: CDK6 knockout HeLa ab260923 cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 37 kDa
Observed band size: 37 kDa
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img137149.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
![Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--immunocytochemistry-immunofluorescence-img69963.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
![Flow Cytometry (Intracellular) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--flow-cytometry-intracellular-img136862.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
![Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395), expandable thumbnail](https://content.abcam.com/products/images/cdk6-antibody-epr4515-bsa-and-azide-free-ab222395--western-blot-img433949.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Cdk6 antibody [EPR4515] - BSA and Azide free (ab222395)")
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