Anti-PKC gamma (phospho T674) antibody (ab5797)
Key features and details
- Rabbit polyclonal to PKC gamma (phospho T674)
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-PKC gamma (phospho T674) antibody
See all PKC gamma primary antibodies -
Description
Rabbit polyclonal to PKC gamma (phospho T674) -
Host species
Rabbit -
Specificity
PKC 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 Applications & Species
See all applications and species dataApplication Species WB Human
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Immunogen
Synthetic phosphopeptide derived from a region of human PKC gamma that contains threonine 674.
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Positive control
- HeLa cells treated with PMA, a phorbol ester.
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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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.3
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA -
Concentration information loading... -
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 gamma. The final product is generated by affinity chromatography using a PKC gamma-derived peptide that is phosphorylated at threonine 674. -
Primary antibody 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. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-PKC gamma (phospho T674) antibody (ab5797)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
µ g/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 pept -
Western blot - Anti-PKC gamma (phospho T674) antibody (ab5797)WB from review by Todd O'Buckley submitted 9 April 2004.
