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Purified bovine brain protein kinase C
Use: It should be possible to selectively block degradation of PKC without affecting the membrane associated activation. This will allow an assessment of the role of proteolysis in the activation of the protein kinase pathway.
Our Abpromise guarantee covers the use of ab31 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.|
|ICC||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|RIA||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.|
|Flow Cyt||Use 1µg for 106 cells.|
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 18368118|
|IHC-P||Use at an assay dependent concentration. PubMed: 17266784|
WB using ab31. Lane 1. Calpain treated PKC from Sigma . Lane 2. Homogenate from rat adipocytes. Lane 3. Homogenate from 3T3-L1 cells (mouse). Lane 4. Purified microsomes from 3T3-L1 cells. Review by Mikael Rutberg submitted 2 September 2004.
Overlay histogram showing Jurkat cells stained with ab31 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab31, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse Alexa Fluor® 488 (IgG; H+L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HL60 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Vero cells with ab31 at a 1/50 dilution. This picture was kindly submitted as part of the review by Martha Simpson-Holley.
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