Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody (ab5817)

Overview

• Product nameAnti-PKC beta 1 + PKC beta 2 (phospho T500) antibodySee all PKC beta 1 + PKC beta 2 primary antibodies ...
• Description
  Rabbit polyclonal to PKC beta 1 + PKC beta 2 (phospho T500)
• SpecificityThis 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 applicationsICC/IF, WB more details
• Species reactivity
  Reacts with: Rat, Human
  Predicted to work with: Mouse
• Immunogen

  Synthetic phosphopeptide derived from a region of human PKC beta 1 & 2 that contains threonine 500.

• Positive controlK562 cells treated with PMA, a phorbol ester.
• 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.

Properties

• FormLiquid
• Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.05% Sodium Azide
  Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3
• Concentration information loading...
• PurityImmunogen affinity purified
• Purification notesThe 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
• IsotypeIgG
• Research Areas
  • Signal Transduction
  • Protein Phosphorylation
  • Ser / Thr Kinases
  • PKC

Applications

Target

• 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.
• Cellular localizationCytoplasm. Nucleus. Membrane.

Target information above from: UniProt accession P05771 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 5579 Human
  • Entrez Gene: 18751 Mouse
  • Entrez Gene: 25023 Rat
  • Omim: 176970 Human
  • SwissProt: P05771 Human
  • SwissProt: P68404 Mouse
  • SwissProt: P68403 Rat
  • Unigene: 460355 Human

  • Unigene: 207496 Mouse
  • Unigene: 446371 Mouse
  • Unigene: 91118 Rat

  see all

• Alternative names
  KPCB_HUMAN antibodyPKC beta antibodyPKC-B antibody

  PKC-beta antibodyPKCB antibodyPKCB1 antibodyPKCB2 antibodyPrkcb antibodyPRKCB I antibodyPRKCB II antibodyPRKCB1 antibodyPRKCB2 antibodyProtein kinase C beta 1 antibodyProtein kinase C beta 2 antibodyProtein kinase C beta antibodyProtein kinase C beta type antibody

  see all

Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody images

• 

  Western blot - Anti-PKC beta 1 + PKC beta 2 (phospho T500) antibody (ab5817)
  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.