Recombinant Anti-CDK5 antibody [EP715Y] - BSA and Azide free (ab220214)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP715Y] to CDK5 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CDK5 antibody [EP715Y] - BSA and Azide free
See all CDK5 primary antibodies -
Description
Rabbit monoclonal [EP715Y] to CDK5 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate, NIH3T3 whole cell lysate, PC-12 whole cell lysate IP: ab220214 + SH-SY5Y whole cell lysate. ICC/IF: SH-SY5Y (Human neuroblastoma epithelial cell)
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General notes
ab220214 is the carrier-free version of ab40773.
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.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP715Y -
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 ab220214 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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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. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa).
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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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. |
WB
Use at an assay dependent concentration. Detects a band of approximately 33 kDa (predicted molecular weight: 33 kDa). |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Probably involved in the control of the cell cycle. Interacts with D1 and D3-type G1 cyclins. Can phosphorylate histone H1, tau, MAP2 and NF-H and NF-M. Also interacts with p35 which activates the kinase. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.
Contains 1 protein kinase domain. -
Cellular localization
Cytoplasm. Cell projection > lamellipodium. Cell projection > growth cone. In axonal growth cone with extension to the peripheral lamellipodia. - Information by UniProt
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Database links
- Entrez Gene: 100190948 Chicken
- Entrez Gene: 1020 Human
- Entrez Gene: 12568 Mouse
- Entrez Gene: 140908 Rat
- Omim: 123831 Human
- SwissProt: Q00535 Human
- SwissProt: P49615 Mouse
- SwissProt: Q03114 Rat
see all -
Alternative names
- Cdk 5 antibody
- Cdk5 antibody
- CDK5_HUMAN antibody
see all
Images
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This data was developed using ab220214, the same antibody clone in a different buffer formulation.
Purified ab220214 at 1:20 dilution (1µg) immunoprecipitating CDK5 in SH-SY5Y whole cell lysate.
Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10µg.
Lane 2 (+): ab220214 + SH-SY5Y whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40773 in SH-SY5Y whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)™(ab131366) (1:1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 33 kDa -
This data was developed using ab220214, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CDK5 with Purified ab220214 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor™ 488, ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue). -
All lanes : Anti-CDK5 antibody [EP715Y] (ab40773) at 1/2000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
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: 33 kDa
Observed band size: 33 kDaBlocking and dilution buffer: 5% NFDM/TBST.
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This data was developed using ab40773, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling CDK5 with Purified ab40773 at 1/50 dilution (4.6 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control. -
All lanes : Anti-CDK5 antibody [EP715Y] (ab40773) at 1/2000 dilution (Unpurified)
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CDK5 knockout HAP1 whole cell lysate
Lane 3 : SHSY5Y whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 33 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40773).
Lanes 1 - 4: Merged signal (red and green). Green - ab40773 observed at 33 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab40773 was shown to specifically react with Cdk5 in wild-type HAP1 cells as signal was lost in CDK5 knockout cells. Wild-type and CDK5 knockout samples were subjected to SDS-PAGE. Ab40773 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (7)
ab220214 has been referenced in 7 publications.
- Kapora E et al. MicroRNA-505-5p functions as a tumor suppressor by targeting cyclin-dependent kinase 5 in cervical cancer. Biosci Rep 39:N/A (2019). PubMed: 31266812
- Yu HP et al. TIGAR regulates DNA damage and repair through pentosephosphate pathway and Cdk5-ATM pathway. Sci Rep 5:9853 (2015). WB . PubMed: 25928429
- Sadleir KR & Vassar R Cdk5 Protein Inhibition and Aß42 Increase BACE1 Protein Level in Primary Neurons by a Post-transcriptional Mechanism: IMPLICATIONS OF CDK5 AS A THERAPEUTIC TARGET FOR ALZHEIMER DISEASE. J Biol Chem 287:7224-35 (2012). WB ; Mouse . PubMed: 22223639
- Zhang HH et al. Increased synaptophysin is involved in inflammation-induced heat hyperalgesia mediated by cyclin-dependent kinase 5 in rats. PLoS One 7:e46666 (2012). IHC-FoFr ; Rat . PubMed: 23056393
- Abdullah C et al. Expression analysis and molecular targeting of cyclin-dependent kinases in advanced melanoma. Cell Cycle 10:977-88 (2011). WB, IHC-Fr ; Human . PubMed: 21358262
- Eggers JP et al. Cyclin-dependent kinase 5 is amplified and overexpressed in pancreatic cancer and activated by mutant K-Ras. Clin Cancer Res 17:6140-50 (2011). IHC-P ; Human . PubMed: 21825040
- Coras R et al. Low proliferation and differentiation capacities of adult hippocampal stem cells correlate with memory dysfunction in humans. Brain 133:3359-72 (2010). PubMed: 20719879