Anti-CDK1 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15)
- Suitable for: WB, IHC-P, ELISA, Dot blot
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-CDK1 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] -
Description
Rabbit monoclonal [EPR7875] to CDK1 + CDK2 + CDK3 + 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 & Species
See all applications and species dataApplication Species IHC-P HumanWB MouseRatHuman -
Immunogen
(phospho Y15). The exact sequence is proprietary.
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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.
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.
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
Images
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Anti-CDK1 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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 + CDK2 + CDK3 + CDK5 (phospho Y15) antibody [EPR7875] (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
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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.
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