Recombinant Anti-PDPK1 antibody [EP569Y] (ab52893)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP569Y] to PDPK1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human, Pig
Related conjugates and formulations
Overview
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Product name
Anti-PDPK1 antibody [EP569Y]
See all PDPK1 primary antibodies -
Description
Rabbit monoclonal [EP569Y] to PDPK1 -
Host species
Rabbit -
Specificity
This antibody detects PDK1 both when phosphorylated and non-phosphorylated on Tyrosine 373 and 376.
The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig.
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Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human, Pig -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC-12, Rat testis, HEK-293, 22Rv1, PIEC, HeLa, K-562, and F9 lysates. IHC-P: human lung carcinoma and human prostate 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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
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
EP569Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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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 ab52893 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 |
1/1000. Detects a band of approximately 64 kDa (predicted molecular weight: 63 kDa).
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IHC-P |
1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig. |
Notes |
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WB
1/1000. Detects a band of approximately 64 kDa (predicted molecular weight: 63 kDa). |
IHC-P
1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig. |
Target
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Function
Phosphorylates and activates not only PKB/AKT, but also PKA, PKC-zeta, RPS6KA1 and RPS6KB1. May play a general role in signaling processes and in development (By similarity). Isoform 3 is catalytically inactive. -
Tissue specificity
Appears to be expressed ubiquitously. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PDK1 subfamily.
Contains 1 PH domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated on tyrosine and serine/threonine. Phosphorylation on Ser-241 in the activation loop is required for full activity. PDK1 itself can autophosphorylate Ser-241, leading to its own activation. -
Cellular localization
Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. Tyrosine phosphorylation seems to occur only at the plasma membrane. - Information by UniProt
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Database links
- Entrez Gene: 5170 Human
- Entrez Gene: 18607 Mouse
- Entrez Gene: 100192445 Pig
- Entrez Gene: 81745 Rat
- Omim: 605213 Human
- SwissProt: O15530 Human
- SwissProt: Q9Z2A0 Mouse
- SwissProt: O55173 Rat
see all -
Alternative names
- 3 phosphoinositide dependent protein kinase 1 antibody
- 3-phosphoinositide-dependent protein kinase 1 antibody
- hPDK 1 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling PDPK1 with Purified ab52893 at 1:150 dilution(2.4 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: PDPK1 knockout HAP1 cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52893 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.ab52893 was shown to recognize PDPK1 when PDPK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and PDPK1 knockout samples were subjected to SDS-PAGE. ab52893 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified) + F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
Predicted band size: 63 kDa
Observed band size: 52-65 kDa why is the actual band size different from the predicted?The molecular weight observed is consistent with what has been described in PMID: 30171166
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All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution
Predicted band size: 63 kDa
Observed band size: 52-65 kDa why is the actual band size different from the predicted?The molecular weight observed is consistent with what has been described in PMID: 30171166
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All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)
Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 2 : Rat testis lysates
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 5 : PIEC (PIG iliac artery endothelial cell) whole cell lysates
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: 63 kDa
Observed band size: 52-65 kDa why is the actual band size different from the predicted?The molecular weight observed is consistent with what has been described in PMID: 30171166
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Ab52893 (1:100) staining human PDPK1 in human prostate carcinoma tissue by immunohistochemistry using paraffin embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution + 293 cell lysate
Predicted band size: 63 kDa
Observed band size: 63 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (25)
ab52893 has been referenced in 25 publications.
- Wu S et al. Telomerase RNA TERC and the PI3K-AKT pathway form a positive feedback loop to regulate cell proliferation independent of telomerase activity. Nucleic Acids Res 50:3764-3776 (2022). PubMed: 35323972
- Fu J et al. MicroRNA-451a prevents cutaneous squamous cell carcinoma progression via the 3-phosphoinositide-dependent protein kinase-1-mediated PI3K/AKT signaling pathway. Exp Ther Med 21:116 (2021). PubMed: 33335579
- Guarnaccia AD et al. Impact of WIN site inhibitor on the WDR5 interactome. Cell Rep 34:108636 (2021). PubMed: 33472061
- Tang N et al. Possible involvement of the Hedgehog and PDPK1-Akt pathways in the growth and migration of small-cell lung cancer. J Int Med Res 49:3000605211016562 (2021). PubMed: 34038205
- Jiang Q et al. SPOP-mediated ubiquitination and degradation of PDK1 suppresses AKT kinase activity and oncogenic functions. Mol Cancer 20:100 (2021). PubMed: 34353330