The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 72, 75 kDa (predicted molecular weight: 72 kDa).
Serine/threonine protein kinase involved in regulating M phase functions during the cell cycle. May also be part of the signaling network controlling cellular adhesion. In vitro, is able to phosphorylate CDC25C and casein.
Transcripts are highly detected in placenta, lung, followed by skeletal muscle, heart, pancreas, ovaries and kidney and weakly detected in liver and brain. May have a short half-live. In cells of hematopoietic origin, strongly and exclusively detected in terminally differentiated macrophages. Transcript expression appears to be down-regulated in primary lung tumor.
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily. Contains 2 POLO box domains. Contains 1 protein kinase domain.
Phosphorylated as cells enter mitosis and dephosphorylated as cells exit mitosis.
All lanes : Anti-PLK3 antibody (ab33119) at 1 µg/ml
Lane 1 : Irradiated Hela Whole Cell Lysate Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 72 kDa Observed band size: 72 kDa Additional bands at: 22 kDa, 38 kDa, 75 kDa (possible post-translational modification). We are unsure as to the identity of these extra bands.
Exposure time: 10 minutes
We hypothesize that the 72 and 75 kDa bands represent the unmodified and phosphorylated forms of PLK3, respectively. This banding pattern of PLK3 is also observed in Bahassi et al., 2004. Oncogene 23, 2658–2663.
ICC/IF image of ab33119 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33119, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Biehs R et al. DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination. Mol Cell65:671-684.e5 (2017).
Read more (PubMed: 28132842) »
Helmke C et al. Ligand stimulation of CD95 induces activation of Plk3 followed by phosphorylation of caspase-8. Cell Res26:914-34 (2016).
Read more (PubMed: 27325299) »