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Purified bovine brain protein kinase C
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.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IHC-FoFr||Use at an assay dependent concentration. PubMed: 18368118|
|IHC-P||Use at an assay dependent concentration. PubMed: 17266784|
ab31 staining PKC in human skin papilloma tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at room temperature; antigen retrieval was by heat mediation in citrate buffer. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. A biotin-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody.
ab31 staining PKC in mouse spleen and duodenum tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/500 in PBS) for 8 hours at 4°C. An Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal (1/1000) was used as the secondary antibody. Nuclei were stained by DAPI.
Review by Mikael Rutberg submitted 2 September 2004.
Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab31 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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, 1 μg/1x106 cells) for 30 minutes 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 minutes at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1 μg/1x106 cells) used under the same conditions. Unlabeled sample (blue line) was 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 HL-60 (Human promyelocytic leukemia cell line) cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.
Vero cells with ab31 at a 1/50 dilution.
HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells with ab31 at 1/50 dilution.
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