Recombinant Anti-PKN antibody [EPR3237] - BSA and Azide free (ab247729)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3237] to PKN - BSA and Azide free
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-PKN antibody [EPR3237] - BSA and Azide free
See all PKN primary antibodies -
Description
Rabbit monoclonal [EPR3237] to PKN - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293T, SH-SY5Y, Jurkat and Human spleen lysates. IHC-P: Human brain tissue.
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General notes
ab247729 is the carrier-free version of ab108976.
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
EPR3237 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab247729 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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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 104 kDa).
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Notes |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Antigen retrieval is recommended.
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WB
Use at an assay dependent concentration. Detects a band of approximately 120 kDa (predicted molecular weight: 104 kDa). |
Target
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Relevance
PKN belongs to the protein kinase C superfamily. This kinase is activated by Rho family of small G proteins and may mediate the Rho-dependent signaling pathway. This kinase can be activated by phospholipids and by limited proteolysis. The 3-phosphoinositide dependent protein kinase-1 (PDPK1/PDK1) is reported to phosphorylate this kinase, which may mediate insulin signals to the actin cytoskeleton. The proteolytic activation of this kinase by caspase-3 or related proteases during apoptosis suggests its role in signal transduction related to apoptosis. -
Cellular localization
Cytoplasmic -
Database links
- Entrez Gene: 5585 Human
- Entrez Gene: 320795 Mouse
- Entrez Gene: 29355 Rat
- SwissProt: Q16512 Human
- SwissProt: P70268 Mouse
- SwissProt: Q63433 Rat
- SwissProt: Q6P748 Rat
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Alternative names
- DBK antibody
- DPKN antibody
- PAK 1 antibody
see all
Images
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All lanes : Anti-PKN antibody [EPR3237] (ab108976) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : PKN1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 104 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?This data was developed using ab108976, the same antibody clone in a different buffer formulation.
Lanes 1-4: Merged signal (red and green). Green - ab108976 observed at 120 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108976 Anti-PKN antibody [EPR3237] was shown to specifically react with PKN in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266599 (knockout cell lysate ab258586) was used. Wild-type and PKN knockout samples were subjected to SDS-PAGE. ab108976 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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This data was developed using ab108976, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of PKN in paraffin embedded Human brain tissue, using ab108976 at a 1/100 dilution. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-PKN antibody [EPR3237] (ab108976) at 1/1000 dilution
Lane 1 : Jurkat lysates
Lane 2 : Human spleen lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 104 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?This data was developed using ab108976, the same antibody clone in a different buffer formulation.
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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Datasheet download
Certificate of Compliance
References (0)
ab247729 has not yet been referenced specifically in any publications.