Anti-PKC iota (phospho T555 + T563) antibody (ab5813)
Key features and details
- Rabbit polyclonal to PKC iota (phospho T555 + T563)
- Suitable for: Flow Cyt, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-PKC iota (phospho T555 + T563) antibody -
Description
Rabbit polyclonal to PKC iota (phospho T555 + T563) -
Host species
Rabbit -
Specificity
This antibody reacts with PKC lambda immunoprecipitates, indicating cross-reactivity for PKC lambda [pT563]. PKC zeta [pT560] (83% homologous) has been shown to cross-react by peptide competition. Peptide competition also suggests that this antibody may partially cross-react with PKC beta 1 [pS642] (58% homologous) and PKC nu [pT655] (42% homologous). -
Tested applications
Suitable for: Flow Cyt, WBmore details -
Species reactivity
Reacts with: Mouse, Human -
Immunogen
Synthetic peptide corresponding to PKC iota (phospho T555 + T563).
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Positive control
- WB: Jurkat cells.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated PKC iota. The final product is generated by affinity chromatography using a PKC iota-derived peptide that is phosphorylated at threonine 555. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5813 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 |
Use at an assay dependent concentration.
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WB |
1/1000. Detects a band of approximately 76 kDa.
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Notes |
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Flow Cyt
Use at an assay dependent concentration. |
WB
1/1000. Detects a band of approximately 76 kDa. |
Target
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Function
Calcium-independent, phospholipid-dependent, serine- and threonine-specific kinase. May play a role in the secretory response to nutrients. Involved in cell polarization processes and the formation of epithelial tight junctions. Implicated in the activation of several signaling pathways including Ras, c-Src and NF-kappa-B pathways. Functions in both pro- and anti-apoptotic pathways. Functions in the RAC1/ERK signaling required for transformed growth. Plays a role in microtubule dynamics through interaction with RAB2A and GAPDH and recruitment to vesicular tubular clusters (VTCs). -
Tissue specificity
Predominantly expressed in lung and brain, but also expressed at lower levels in many tissues including pancreatic islets. Highly expressed in non-small cell lung cancers. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 OPR domain.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain. -
Domain
The OPR domain mediates interaction with SQSTM1.
The C1 domain does not bind diacylglycerol (DAG). -
Post-translational
modificationsOn neuronal growth factor (NGF) stimulation, phosphorylated by Src on Tyr-265, Tyr-280 and Tyr-334. Phosphorylation on Tyr-265 facilitates binding to KPNB1/importin-beta regulating entry of PRKCI into the nucleus. Phosphorylation on Tyr-334 is important for NF-kappa-B stimulation. -
Cellular localization
Cytoplasm. Membrane. Endosome. Nucleus. Transported into the endosome through interaction with SQSTM1/p62. After phosphorylation by cSrc, transported into the nucleus through interaction with KPNB1. Colocalizes with CDK7 in the cytoplasm and nucleus. Vesicular tubular clusters. Transported to VTCs through interaction with RAB2A. - Information by UniProt
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Database links
- Entrez Gene: 5584 Human
- Entrez Gene: 18759 Mouse
- Omim: 600539 Human
- SwissProt: P41743 Human
- SwissProt: Q62074 Mouse
- Unigene: 478199 Human
- Unigene: 291554 Mouse
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Alternative names
- aPKC lambda/iota antibody
- aPKC-lambda/iota antibody
- Atypical protein kinase C lambda/iota antibody
see all
Images
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Peptide Competition and Phosphatase Treatment: Lysates prepared from Jurkat cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% low-fat milk-TBST buffer for one hour at room temperature, and incubated with ab5813 antibody for two hours at room temperature in a 3% low-fat milk-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that the phosphopeptide corresponding to PKC iota [pT555] blocks the antibody signal. The peptide corresponding to PKC zeta [pT560] blocks the antibody signal and the peptides corresponding to PKC isoforms beta 1 [pT642] and gamma [pÔ655] partially block the antibody signal (data not shown), suggesting cross-reactivity of the antibody with these sites. The antibody signal was not blocked by the corresponding peptides of any other PKC isoforms. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
Peptide Competition and Phosphatase Treatment: Lysates prepared from Jurkat cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-4) or treated with lambda phosphatase (5), blocked with a 5% low-fat milk-TBST buffer for one hour at room temperature, and incubated with ab5813 antibody for two hours at room temperature in a 3% low-fat milk-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignalTM method. The data show that the phosphopeptide corresponding to PKC iota [pT555] blocks the antibody signal. The peptide corresponding to PKC zeta [pT560] blocks the antibody signal and the peptides corresponding to PKC isoforms beta 1 [pT642] and gamma [pÔ655] partially block the antibody signal (data not shown), suggesting cross-reactivity of the antibody with these sites. The antibody signal was not blocked by the corresponding peptides of any other PKC isoforms. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
Datasheets and documents
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SDS download
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Datasheet download
References (20)
ab5813 has been referenced in 20 publications.
- Dey A et al. The interruption of atypical PKC signaling and Temozolomide combination therapy against glioblastoma. Cell Signal 77:109819 (2021). PubMed: 33147518
- Patel R et al. Simultaneous inhibition of atypical protein kinase-C and mTOR impedes bladder cancer cell progression. Int J Oncol 56:1373-1386 (2020). PubMed: 32236625
- Kotula-Balak M et al. Towards understanding leydigioma: do G protein-coupled estrogen receptor and peroxisome proliferator-activated receptor regulate lipid metabolism and steroidogenesis in Leydig cell tumors? Protoplasma 257:1149-1163 (2020). PubMed: 32180008
- Ito M et al. Targeting PKC?-PAK1 in EGFR-mutation positive non-small cell lung cancer. Transl Lung Cancer Res 8:667-673 (2019). PubMed: 31737502
- Li HS et al. Protein Kinase C Lambda Mediates Acid-Sensing Ion Channel 1a-Dependent Cortical Synaptic Plasticity and Pain Hypersensitivity. J Neurosci 39:5773-5793 (2019). PubMed: 31101759