Anti-PKC gamma (phospho T674) antibody (ab5797)

Overview

  • Product nameAnti-PKC gamma (phospho T674) antibody
    See all PKC gamma primary antibodies
  • Description
    Rabbit polyclonal to PKC gamma (phospho T674)
  • SpecificityPKC alpha (69%) may cross-react in cells expressing a high level of this protein. The peptide competition data (see Figure) suggest that this antibody will cross-react with PKC alpha [pS657] (69% homologous), beta 1 [pS661] (54%) and PKC eta [pS674] (38%).
  • Tested applicationsSuitable for: WBmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic phosphopeptide derived from a region of human PKC gamma that contains threonine 674.

  • Positive control
    • HeLa cells treated with PMA, a phorbol ester.
  • General notes


    Protein Kinase C gamma (PKC gamma) is an 80 kDa member of the conventional grou(cPKCs: sensitive to calcium, 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 and transcriptional regulation. PKC gamma plays a key role in neuronal signal transduction and in regulating intercellular communication. The activation loop threonine (threonine 514 in PKC gamma) of conventional PKCs is phosphorylated by phosphoinositide-dependent kinase-1 (PDK1), which is necessary for its autophosphorylation on threonine 655 in the turn loop, and threonine 674 in the hydrophobic loop of the carboxy terminus, a critical step in generating a catalytically mature enzyme. The phosphorylation of the hydrophobic loop in the carboxyl terminus of PKCs is believed to be a key determinant in regulating PKC interaction with PDK1.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPreservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA
  • 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 gamma. The final product is generated by affinity chromatography using a PKC gamma-derived peptide that is phosphorylated at threonine 674.
  • Primary antibody notesProtein Kinase C gamma (PKC gamma) is an 80 kDa member of the conventional grou(cPKCs: sensitive to calcium, 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 and transcriptional regulation. PKC gamma plays a key role in neuronal signal transduction and in regulating intercellular communication. The activation loop threonine (threonine 514 in PKC gamma) of conventional PKCs is phosphorylated by phosphoinositide-dependent kinase-1 (PDK1), which is necessary for its autophosphorylation on threonine 655 in the turn loop, and threonine 674 in the hydrophobic loop of the carboxy terminus, a critical step in generating a catalytically mature enzyme. The phosphorylation of the hydrophobic loop in the carboxyl terminus of PKCs is believed to be a key determinant in regulating PKC interaction with PDK1.
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5797 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
WB 1/1000. Detects a band of approximately 80 kDa.

Target

  • FunctionThis is a calcium-activated, phospholipid-dependent, serine- and threonine-specific enzyme.
    PKC is activated by diacylglycerol which in turn phosphorylates a range of cellular proteins. PKC also serves as the receptor for phorbol esters, a class of tumor promoters.
  • Tissue specificityExpressed in Purkinje cells of the cerebellar cortex.
  • Involvement in diseaseDefects in PRKCG are the cause of spinocerebellar ataxia type 14 (SCA14) [MIM:605361]. Spinocerebellar ataxia is a clinically and genetically heterogeneous group of cerebellar disorders. Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements, due to degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA14 is an autosomal dominant cerebellar ataxia (ADCA).
  • 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • KPCG_HUMAN antibody
    • MGC57564 antibody
    • OTTHUMP00000067291 antibody
    • PKC-gamma antibody
    • PKCC antibody
    • PKCG antibody
    • PRKCG antibody
    • Protein kinase C gamma antibody
    • Protein kinase C gamma polypeptide antibody
    • Protein kinase C gamma type antibody
    • Protein kinase C, gamma antibody
    • SCA 14 antibody
    • SCA14 antibody
    see all

Anti-PKC gamma (phospho T674) antibody images

  • Peptide Competition and Phosphatase Treatment: Lysates prepared from HeLa cells stimulated with PMA were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-13) or treated with lambda (ë) phosphatase (14), blocked with a 5% BSA-TBST buffer overnight at 4°C, and incubated with 0.50 µg/mL ab5797 antibody for two hours at room temperature in a 3% BSA TBST buffer, following prior incubation with: the phosphopeptide corresponding to the immunogen from other PKC isoforms (1-9), the phosphopeptide immunogen (10), the non-phosphopeptide corresponding to the immunogen (11), a generic phosphothreonine containing peptide (12) or, no peptide (13, 14). 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 peptide corresponding to PKC gamma [pT674] blocks the antibody signal. The peptides corresponding to PKC alpha [pT657], beta 1 [pS661] and PKC eta [pS674] also block the antibody signal, suggesting cross-reactivity of ab5797 antibody with these sites. The antibody signal was not blocked by the peptides corresponding to PKC isoforms beta 2 [pT660], gamma [pT514], gamma [pT655], delta [pS664], theta [pS695] and zeta [pT560], thereby demonstrating the specificity of the antibody. The data also show that phosphatase stripping eliminates the signal, verifying that the antibody is phospho-specific.
  • WB from review by Todd O'Buckley submitted 9 April 2004.

References for Anti-PKC gamma (phospho T674) antibody (ab5797)

This product has been referenced in:
  • Schmidt HD  et al. Stimulation of mGluR5 in the accumbens shell promotes cocaine seeking by activating PKC gamma. J Neurosci 33:14160-9 (2013). Read more (PubMed: 23986250) »

See 1 Publication for this product

Product Wall


HeLa cells were treated with 1 µM PMA for 30 minutes for the western blot image of ab5797.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - other (Rat Brain, pooled nucleus accumbens)
Specification Rat Brain, pooled nucleus accumbens
Blocking step Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1%
Username

Dr. Stephen Kelley

Verified customer

Submitted Sep 02 2005

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