Product nameAnti-PKC delta (phospho S645) antibody [EP1486Y]
See all PKC delta primary antibodies
DescriptionRabbit monoclonal [EP1486Y] to PKC delta (phospho S645)
Tested applicationsSuitable for: ELISA, Dot blot, WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt or IP
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human PKC delta aa 600 to the C-terminus (phospho S645).
Database link: Q05655
- 293T cell lysate; Human breat carcinoma; HeLa cells
A trial size is available to purchase for this antibody.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab108972 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|WB||1/10000 - 1/50000. Predicted molecular weight: 78 kDa.|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Antigen retrieval is recommended.
|ICC/IF||1/50 - 1/100.|
FunctionThis is calcium-independent, 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. May play a role in antigen-dependent control of B-cell function. Phosphorylates MUC1 in the C-terminal and regulates the interaction between MUC1 and beta-catenin.
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.
DomainThe C1 domain, containing the phorbol ester/DAG-type region 1 (C1A) and 2 (C1B), is the diacylglycerol sensor.
The C2 domain is a non-calcium binding domain. It binds proteins containing phosphotyrosine in a sequence-specific manner.
modificationsPhosphorylated on Thr-507, within the activation loop. Autophosphorylated and/or phosphorylated. Although the Thr-507 phosphorylation occurs it is not a prerequisite for enzymatic activity.
Cellular localizationCytoplasm. Membrane.
- Information by UniProt
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Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
Lanes 2, 6 and 10: PKC delta knockout HAP1 cell lysate (20 µg)
Lanes 3, 7 and 11: A431 cell lysate (20 µg)
Lanes 4, 8 and 12: HeLa cell lysate (20 µg)
Lanes 1, 2, 3 and 4: Green signal from target – ab108972 observed at 78 kDa
Lanes 5, 6, 7 and 8: Red signal from loading control – ab8245 observed at 37 kDa
Lanes 9, 10, 11 and 12: Merged (red and green) signal
ab108972 was shown to specifically react with PKC delta when PKC delta knockout samples were used. Wild-type and PKC delta knockout samples were subjected to SDS-PAGE. ab108972 and ab8245 (loading control to GAPDH) were diluted 1/10000 and 1/2000 respectively and incubated overnight at 4ºC. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.
Immunofluorescent staining of PKC delta in HeLa cells, using ab108972 at a 1/50 dilution.
Immunohistochemical analysis of PKC delta in paraffin embedded Human breast carcinoma tissue, using ab108972 at a 1/50 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1:20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: THP-1 whole cell lysate treated with Calyculin A with no peptides, Lane 2: THP-1 whole cell lysate treated with Calyculin A with PKCd S645Pho peptides, Lane 3: THP-1 whole cell lysate treated with Calyculin A with PKCd unmodified peptides, Lane 4: None, Observed MW 78 kDa, Exposure time 10 seconds
Primary ab Dilution 1:100,000 dilution, Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1: 20,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM/TBST, Lane 1: Untreated THP-1 (Human monocytic leukemia cell line) whole cell lysates 15ug, Lane 2: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes whole cell lysates 15ug. Lane 3: THP-1 (Human monocytic leukemia cell line) treated with 50nM Calyculin A for 60 minutes’ whole cell lysates 15ug. Then the membrane was incubated with phosphatase. Lane 4: None, Observed MW 78 kDa, Exposure time 5 seconds
Primary ab dilution 1: 1000 dilution (2.355 μg /ml), Secondary ab description and code (ab id) Goat Anti-Rabbit IgG, (H+L), HRP conjugated (ab97051), Secondary ab dilution 1:100,000 dilution, Blocking buffer and concentration 5% NFDM/TBST, Diluting buffer and concentration 5% NFDM /TBST, Lane 1: PKCd S645Pho peptides, Lane 2: PKCd unmodified peptides, Lane 3: None, Lane 4: None, Observed MW N/A, Exposure time 3 minutes
Antigen PKCd S645Pho peptide, PKCd S645Pho unmodified peptide, Antigen concentration 1000ng/ml, Primary antibody concentration range 0~1000ng/ml, Secondary antibody Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Secondary antibody concentration 1:2500 dilution