Recombinant Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP435Y-129R] to CDKN2A/p16INK4A +CDKN2B/p15INK4B - BSA and Azide free
- Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free
See all CDKN2A/p16INK4A +CDKN2B/p15INK4B primary antibodies -
Description
Rabbit monoclonal [EP435Y-129R] to CDKN2A/p16INK4A +CDKN2B/p15INK4B - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, Flow Cyt (Intra), WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK-293, HeLa ICC/IF: HeLa cells Flow Cyt (intra): HEK-293 cells. IP: HEK-293
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General notes
ab219723 is the carrier-free version of ab81278.
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.
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. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP435Y-129R -
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 ab219723 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 |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 17 kDa.
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IP |
Use at an assay dependent concentration.
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Notes |
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ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 17 kDa. |
IP
Use at an assay dependent concentration. |
Target
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Function
Acts as a negative regulator of the proliferation of normal cells by interacting strongly with CDK4 and CDK6. This inhibits their ability to interact with cyclins D and to phosphorylate the retinoblastoma protein. -
Tissue specificity
Widely expressed but not detected in brain or skeletal muscle. Isoform 3 is pancreas-specific. -
Involvement in disease
The association between cutaneous and uveal melanomas in some families suggests that mutations in CDKN2A may account for a proportion of uveal melanomas. However, CDKN2A mutations are rarely found in uveal melanoma patients.
Melanoma, cutaneous malignant 2
Familial atypical multiple mole melanoma-pancreatic carcinoma syndrome
Melanoma-astrocytoma syndrome -
Sequence similarities
Belongs to the CDKN2 cyclin-dependent kinase inhibitor family.
Contains 4 ANK repeats. -
Post-translational
modificationsPhosphorylation seems to increase interaction with CDK4. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 1029 Human
- SwissProt: P42771 Human
- Unigene: 512599 Human
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Alternative names
- CDK4I antibody
- CDKN2A antibody
- CDN2A_HUMAN antibody
see all
Images
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Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilised HeLa (human cervix adenocarcinoma epithelial cell) labelling CDKN2A/p16INK4a with ab81278 at 1/250 dilution, followed by AlexaFluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HeLa is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain alpha Tubulin at 1/200 dilution (red). The Nuclear counterstain was DAPI (blue).
Secondary antibody only control: Secondary antibody is AlexaFluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (green).
This data was developed using ab81278, the same antibody clone in a different buffer formulation.
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Flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilised HEK-293 (Human embryonic kidney epithelial cell) with ab81278 at 1/20 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
This data was developed using ab81278, the same antibody clone in a different buffer formulation.
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Clone EP435Y-129R (ab219723) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4A +CDKN2B/p15INK4B antibody [EP435Y-129R] (Alexa Fluor® 647). Please refer to ab199819 for protocol details.
ab199819 staining CDKN2A/p16INK4a in HeLa cells. 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 overnight at +4°C with ab199819 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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All lanes : Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723) at 1/10000 dilution
Lane 1 : HEK-293 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 1/20 dilution per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution
Predicted band size: 17 kDa
Exposure time: 3 seconds -
Clone EP435Y-129R (ab219723) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4a antibody [EP435Y-129R] (Alexa Fluor® 488). Please refer to ab199756 for protocol details.
ab199756 staining CDKN2A/p16INK4a in HeLa cells. 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 overnight at +4°C with ab199756 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293 (Human embryonic kidney epithelial cell) cells labelling CDKN2A with ab219723 at 1/100 dilution (1µg)/ Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.
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CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg with ab219723 at 1:50 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab219723 1:1000 dilution (2 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2: ab219723 IP in HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219723 in HEK-293 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab219723 has not yet been referenced specifically in any publications.