The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Detects a band of approximately 80 kDa (predicted molecular weight: 77 kDa).
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.
Post-translational 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.
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: PKC delta knockout HAP1 cell lysate (20 µg) Lane 3: A431 cell lysate (20 µg) Lane 4: HeLa cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab86800 observed at 80 kDa. Red - loading control, ab8245, observed at 37 kDa. ab86800 was shown to recognize PKC delta when PKC delta knockout samples were used, along with additional cross-reactive bands. Wild-type and PKC delta knockout samples were subjected to SDS-PAGE. ab86800 and ab8245 (loading control to GAPDH) were diluted 1 μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - PKC delta antibody (ab86800)
All lanes : Anti-PKC delta antibody (ab86800) at 1 µg/ml
Lane 1 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 2 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution Developed using the ECL technique
Exposure time : 12 minutesHuman Protein kinase C delta type (PKC delta) contains a number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Western blot - PKC delta protein (Active) (ab60844)
Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 77 kDa
Exposure time : 4 minutes
Western blot - Anti-PKC delta antibody (ab86800)
Anti-PKC delta antibody (ab86800) at 1 µg/ml + Brain (Rat) Tissue Lysate at 10 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 77 kDa Observed band size : 77 kDa
Exposure time : 20 minutes
References for Anti-PKC delta antibody (ab86800)
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