Recombinant Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7875] to CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15)
- Suitable for: WB, IHC-P, ELISA, Dot blot
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR -
Description
Rabbit monoclonal [EPR7875] to CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) -
Host species
Rabbit -
Specificity
The antibody will cross-react with phosphorylated CDK1 (pY15), CDK2 (pY15), CDK3 (pY15) and CDK5 (pY15) but not with non-phosphorylated CDK1, CDK2, CDK3 and CDK5. Please see our ELISA results on the images section. -
Tested applications
Suitable for: WB, IHC-P, ELISA, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- HeLa cell lysate treated with UV, human colon, C6 cell lysate, NIH/3T3 cell lysate, Saso2 cells, human breast carcinoma tissue
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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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 4.10 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR7875 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133463 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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WB | (1) |
1/1000 - 1/2000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
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IHC-P | (1) |
1/50 - 1/75. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ELISA |
Use at an assay dependent concentration.
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Dot blot |
1/1000.
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Notes |
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WB
1/1000 - 1/2000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa). |
IHC-P
1/50 - 1/75. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ELISA
Use at an assay dependent concentration. |
Dot blot
1/1000. |
Target
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Cellular localization
CDK1: Nucleus. CDK5: Cytoplasm. Cell projection > lamellipodium. Cell projection > growth cone. In axonal growth cone with extension to the peripheral lamellipodia. -
Database links
- Entrez Gene: 1017 Human
- Entrez Gene: 1018 Human
- Entrez Gene: 1020 Human
- Entrez Gene: 983 Human
- Entrez Gene: 12534 Mouse
- Entrez Gene: 12566 Mouse
- Entrez Gene: 12568 Mouse
- Entrez Gene: 140908 Rat
see all -
Form
CDK1: CDK1 can be located to the Nucleus, cytoplasm and Mithocondria. It's cytoplasmic during interphase and reversibly translocated from cytoplasm to the nucleus when phosphorilated before G2-M transition when associated with cyclin-B1. Accumulates in mitochondria in G2-arrested cells upon DNA-damage.
Images
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Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/1000 dilution (purified) + HeLa cell lysate treated with UV at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemical staining of paraffin embedded human colon with purified ab133463 at a working dilution of 1 in 75. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Direct ELISA antibody dose-response curve using purified ab133463 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
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Direct ELISA antibody dose-response curve using purified ab133463 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
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Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/2000 dilution (purified) + C6 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/2000 dilution (purified) + NIH/3T3 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Dot blot analysis of CDK1+CDK2+CDK3+CDK5 (pY15) phospho peptide (lane 1) and CDK1+CDK2+CDK3+CDK5 non-phospho peptide (lane 2) labelling CDK1+CDK2+CDK3+CDK5 (phospho Y15) with unpurified ab133463 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 10 seconds.
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Saso2 cells were incubated at 37°C for 24 hours with vehicle control (0 μM) and different concentrations of Amifostine (ab141060). Decreased expression of CDK1 (phospho Y15) (unpurified ab133463) in Saso2 cells correlates with an increase in Amifostine concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with unpurified ab133463 at 1 μg/ml and ab32384 at 1 μg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 and visualised using ECL development solution.
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Immunohistochemical analysis of paraffin embedded human breast carcinoma tissue labelling CDK1+CDK2+CDK3+CDK5 with unpurified ab133463 at 1/50 dilution. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/1500 dilution (unpurified) + NIH/3T3 cell lysate at 1/1000 dilution
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/1500 dilution (unpurified) + C6 cell lysate at 10 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-CDK1 (phospho Y15) + CDK2 (phospho Y15) + CDK3 (phospho Y15) + CDK5 (phospho Y15) antibody [EPR (ab133463) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : HeLa cell lysate treated with UV
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa -
Immunohistochemical staining of paraffin embedded human colon with unpurified ab133463 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab133463 has been referenced in 14 publications.
- Gray S et al. Cyclin N-Terminal Domain-Containing-1 Coordinates Meiotic Crossover Formation with Cell-Cycle Progression in a Cyclin-Independent Manner. Cell Rep 32:107858 (2020). PubMed: 32640224
- Kok YP et al. Overexpression of Cyclin E1 or Cdc25A leads to replication stress, mitotic aberrancies, and increased sensitivity to replication checkpoint inhibitors. Oncogenesis 9:88 (2020). PubMed: 33028815
- Corella AN et al. Identification of Therapeutic Vulnerabilities in Small-cell Neuroendocrine Prostate Cancer. Clin Cancer Res 26:1667-1677 (2020). PubMed: 31806643
- Lee JH et al. C/EBPß Is a Transcriptional Regulator of Wee1 at the G2/M Phase of the Cell Cycle. Cells 8:N/A (2019). PubMed: 30754676
- Heijink AM et al. Modeling of Cisplatin-Induced Signaling Dynamics in Triple-Negative Breast Cancer Cells Reveals Mediators of Sensitivity. Cell Rep 28:2345-2357.e5 (2019). PubMed: 31461651