The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/250. Predicted molecular weight: 80 kDa. (AP/HRP). With a more sensitive detection system, such as enhanced chemiluminescence or ELISA, the antibody may be diluted further.
This is a calcium-activated, phospholipid-dependent, serine- and threonine-specific enzyme. May play a role in cell motility by phosphorylating CSPG4. 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.
Belongs 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.
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: PKC alpha knockout HAP1 cell lysate (20 µg) Lane 3: K562 cell lysate (20 µg) Lane 4: HEK293 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab4124 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab4124 was shown to recognize PKC alpha in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when PKC alpha knockout samples were examined. Wild-type and PKC alpha knockout samples were subjected to SDS-PAGE. ab4124 and ab8245 (loading control to GAPDH) were diluted to 1/250 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-PKC alpha antibody (ab4124)
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: PKC alpha knockout HAP1 cell lysate (20 µg) Lane 3: K562 cell lysate (20 µg) Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab4124 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab4124 and a competitor's top cited rabbit polyclonal antibody.
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