Anti-Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) antibody [EPR2233Y] (ab76146)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2233Y] to Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15)
- Suitable for: IHC-P, Dot blot, WB, IP
- Reacts with: Rat, Human
Overview
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Product name
Anti-Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) antibody [EPR2233Y] -
Description
Rabbit monoclonal [EPR2233Y] to Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) -
Host species
Rabbit -
Specificity
ab76146 detects Cdk1, 2, 3, 5 phosphorylated on tyrosine 15.
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Tested Applications & Species
See all applications and species dataApplication Species IHC-P RatHumanIP HumanWB Human -
Immunogen
Synthetic peptide within Human CDK2 (phospho Y15). The exact sequence is proprietary.
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Positive control
- WB: HeLa cell and mouse spleen tissue lysates; 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged blank expression vector, whole cell lysate; 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1/2/3/5 expression vector, whole cell lysate IHC-P: Human transitional cell carcinoma of bladder and rat colon tissues. IP: HeLa whole cell lysate (ab150035).
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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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2233Y -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat colon tissue labelling Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) with purified ab76146 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP polymer-conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) antibody [EPR2233Y] (ab76146) at 1/1000 dilution
Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged blank expression vector, whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1 expression vector, whole cell lysate
Lane 3 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK1 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
Lane 4 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK2 expression vector, whole cell lysate
Lane 5 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK2 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
Lane 6 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK3 expression vector, whole cell lysate
Lane 7 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK3 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
Lane 8 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK5 expression vector, whole cell lysate
Lane 9 : 293T (human embryonic kidney epithelial cell) transfected with GFP-tagged CDK5 expression vector, whole cell lysate. Then the membrane was incubated with phosphatase
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
Observed band size: 34,65 kDa why is the actual band size different from the predicted?ab76146 detects Cdk1, 2, 3, 5 phosphorylated on tyrosine 15.
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All lanes : Anti-Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) antibody [EPR2233Y] (ab76146) at 1/50000 dilution (purified)
Lane 1 : Untreated HeLa cell lysate
Lane 2 : HeLa cell lysate - treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 34 kDa
Exposure time: 10 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Dot blot analysis of Cdk2 (pY15) peptide (Lane 1) and Cdk2 non-phospho peptide (Lane 2) labelling Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) with purified ab76146 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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ab76146 (purified) at 1/100 immunoprecipitating Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) in HeLa cell lysates (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of bladder tissue labelling Cdk1 + Cdk2 + Cdk3 + Cdk5 (phospho Y15) with purified ab76146 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer, pH 9. A prediluted HRP polymer-conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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