The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/50000 - 1/200000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
1/50 - 1/100.
Application notesIs unsuitable for Flow Cyt or IP.
FunctionCalcium-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 similaritiesBelongs 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.
Dot blot analysis of PKC beta 2 (phospho S660) phospho peptide (Lane 1), PKC beta 2 Non-phospho peptide (Lane 2), labelling PKC beta 2 (phospho S660) with ab75837 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/2500.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
References for Anti-PKC beta 2 (phospho S660) antibody [EP1902Y] (ab75837)
Qu C et al. Trimeric G protein-CARMA1 axis links smoothened, the hedgehog receptor transducer, to NF-?B activation in diffuse large B-cell lymphoma. Blood121:4718-28 (2013).
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