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, WBmore 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 control
    • K562 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

Applications

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
ICC/IF
WB
  • Application notesICC/IF: 1/1000. Punctate staining observed.
    WB: 1/1000. Detects a band of approximately 80 kDa.

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • 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
      modifications
      Phosphorylation 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.
    • Information by UniProt
    • Database links
    • Alternative names
      • KPCB_HUMAN antibody
      • PKC beta antibody
      • PKC-B antibody
      • PKC-beta antibody
      • PKCB antibody
      • PKCB1 antibody
      • PKCB2 antibody
      • Prkcb antibody
      • PRKCB I antibody
      • PRKCB II antibody
      • PRKCB1 antibody
      • PRKCB2 antibody
      • Protein kinase C beta 1 antibody
      • Protein kinase C beta 2 antibody
      • Protein kinase C beta antibody
      • Protein kinase C beta type antibody
      see all

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

    • 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.

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

    This product has been referenced in:
    • 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 . Read more (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). Read more (PubMed: 18006785) »

    See all 2 Publications for this product

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Sample Rat Cell (Cerebellar neurons in culture)
    Specification Cerebellar neurons in culture
    Fixative Formaldehyde
    Blocking step Gelatin from fish as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25
    Username

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    Submitted Apr 09 2007

    Application Western blot
    Sample Human Cell lysate - whole cell (BxPC3 cells)
    Loading amount 20 µg
    Specification BxPC3 cells
    Treatment 10% FBS
    Blocking step BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5%
    Username

    Abcam user community

    Verified customer

    Submitted Oct 23 2006

    Further to a follow up email from our source of this antibody I have ascertained that A431 cells are apparently NOT a good choice for the detection of PKC beta. According to them the cells have been shown to express PKC alpha, delta and zeta but not PK...

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    Thank you for getting back to me. I have been unable to obtain a satisfactory answer from the source of this antibody. However, I see no reason why the positive control that you have been using would not be suitable. Furthermore the approach that y...

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    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionaire and I have a few comments. The approach that you are employing is largely one that I would recomme...

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    Thank you for your enquiry. This testing has not been performed as far as I know, but with some research I have found the following information: PKC alpha shows ~79% homology within the range of the immunogen sequence. PKC beta I shows 50% homol...

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    Thank you for your enquiry. Are you interested in detecting PKC beta 1 or 2? Ab5785 detects PKC beta 2 phosphorylated at T641, where as ab5817 detects PKC beta 1 and 2 phosphorylated at T500. For both of these antibodies there are Western blot imag...

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    Thank you for your enquiry. Useful working concentration range of PMA is 1–100 nM. You may need to consider testing the half-maximal effective dose of PMA in your experimental system. Please take a look at the following website, you may find so...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"