Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody (ab5817)
Key features and details
- Rabbit polyclonal to PKC beta 1 + PKC beta 2 (phospho T500)
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody
See all PKC beta 1 + PKC beta 2 primary antibodies -
Description
Rabbit polyclonal to PKC beta 1 + PKC beta 2 (phospho T500) -
Host species
Rabbit -
Specificity
This antibody cross-reacts with PKC alpha [pT497] (88% homologous) and partially cross-reacts with PKC gamma [pT514] (63% homologous) and epsilon [pT566] (75% homologous), as determined by peptide competition experiments. -
Tested applications
Suitable for: WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse -
Immunogen
Synthetic peptide corresponding to PKC beta 1 + PKC beta 2 (phospho T500).
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General notes
Protein Kinase C beta (PKC beta) is an 80 kDa member of the conventional group (cPKCs: sensitive to diacylglycerol, phosphotidylserine and phorbol esters) of the PKC family of serine/threonine kinases that are involved in a wide range of physiological processes including mitogenesis, cell survival, transcriptional regulation and tumor promotion. PKC beta has been implicated in diabetes and carcinogenesis. PKC beta isoforms 1 & 2 are phosphorylated on three sites, threonine 500 in the activation loop, beta 1 threonine 642 (beta 2 641) in the turn loop and beta 1 serine 661 (beta 2 660) in the hydrophobic loop. Phosphorylation of PKC beta 1 & 2 on threonine 500 by PDK1 is a prerequisite for its autophosphoylation and catalytic competence.
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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, 50% Glycerol, 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 beta. The final product is generated by affinity chromatography using a PKC beta-derived peptide that is phosphorylated at threonine 500. -
Primary antibody notes
Protein Kinase C beta (PKC beta) is an 80 kDa member of the conventional group (cPKCs: sensitive to diacylglycerol, phosphotidylserine and phorbol esters) of the PKC family of serine/threonine kinases that are involved in a wide range of physiological processes including mitogenesis, cell survival, transcriptional regulation and tumor promotion. PKC beta has been implicated in diabetes and carcinogenesis. PKC beta isoforms 1 & 2 are phosphorylated on three sites, threonine 500 in the activation loop, beta 1 threonine 642 (beta 2 641) in the turn loop and beta 1 serine 661 (beta 2 660) in the hydrophobic loop. Phosphorylation of PKC beta 1 & 2 on threonine 500 by PDK1 is a prerequisite for its autophosphoylation and catalytic competence. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5817 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) |
1/1000. Detects a band of approximately 80 kDa.
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Notes |
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WB
1/1000. Detects a band of approximately 80 kDa. |
Target
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Function
Calcium-activated and phospholipid-dependent serine/threonine-protein kinase involved in various processes such as regulation of the B-cell receptor (BCR) signalosome, apoptosis and transcription regulation. Plays a key role in B-cell activation and function by regulating BCR-induced NF-kappa-B activation and B-cell suvival. Required for recruitment and activation of the IKK kinase to lipid rafts and mediates phosphorylation of CARD11/CARMA1 at 'Ser-559', 'Ser-644' and 'Ser-652', leading to activate the NF-kappa-B signaling. Involved in apoptosis following oxidative damage: in case of oxidative conditions, specifically phosphorylates 'Ser-36' of isoform p66Shc of SHC1, leading to mitochondrial accumulation of p66Shc, where p66Shc acts as a reactive oxygen species producer. Acts as a coactivator of androgen receptor (ANDR)-dependent transcription, by being recruited to ANDR target genes and specifically mediating phosphorylation of 'Thr-6' of histone H3 (H3T6ph), a specific tag for epigenetic transcriptional activation that prevents demethylation of histone H3 'Lys-4' (H3K4me) by LSD1/KDM1A. Also involved in triglyceride homeostasis. Serves as the receptor for phorbol esters, a class of tumor promoters. -
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 C2 domain.
Contains 2 phorbol-ester/DAG-type zinc fingers.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylation on Thr-500 within the activation loop renders it competent to autophosphorylate. Subsequent autophosphorylation of Thr-642 maintains catalytic competence, and autophosphorylation on Ser-661 appears to release the kinase into the cytosol. Autophosphorylation on other sites i.e. in the N-terminal and hinge regions have no effect on enzyme activity. -
Cellular localization
Cytoplasm. Nucleus. Membrane. - Information by UniProt
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Database links
- Entrez Gene: 5579 Human
- Entrez Gene: 18751 Mouse
- Omim: 176970 Human
- SwissProt: P05771 Human
- SwissProt: P68404 Mouse
- Unigene: 460355 Human
- Unigene: 207496 Mouse
- Unigene: 446371 Mouse
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Alternative names
- KPCB_HUMAN antibody
- PKC beta antibody
- PKC-B antibody
see all
Images
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Peptide Competition and Phosphatase Treatment: Lysates prepared from K562 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% BSA-TBST buffer for one hour at room temperature, and incubated with ab5817 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), 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 only the peptide corresponding to PKC beta 1 & 2 [pT500] blocks the antibody signal. 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 K562 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% BSA-TBST buffer for one hour at room temperature, and incubated with ab5817 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5), 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 only the peptide corresponding to PKC beta 1 & 2 [pT500] blocks the antibody signal. 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 (3)
ab5817 has been referenced in 3 publications.
- Asami M et al. Dopamine and the phosphorylated dopamine transporter are increased in the diacylglycerol kinase ?-knockout mouse brain. FEBS Lett 595:1313-1321 (2021). PubMed: 33599293
- Molè D et al. Protein kinase C: a putative new target for the control of human medullary thyroid carcinoma cell proliferation in vitro. Endocrinology 153:2088-98 (2012). WB ; Human . PubMed: 22374978
- Spalding AC et al. Inhibition of protein kinase Cbeta by enzastaurin enhances radiation cytotoxicity in pancreatic cancer. Clin Cancer Res 13:6827-33 (2007). PubMed: 18006785