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Our Abpromise guarantee covers the use of ab73406 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 85 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Cytosolic Phospholipase A2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: A549 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab73406 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab73406 was shown to specifically recognize Cytosolic Phospholipase A2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Cytosolic Phospholipase A2 knockout samples were examined. Wild-type and Cytosolic Phospholipase A2 knockout samples were subjected to SDS-PAGE. Ab73406 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
The 100kDa band is comparable to molecular weights seen with other commercially available antibodies to Cytosolic Phospholipase A2
IHC image of Cytosolic Phospholipsae A2 staining in humam colon carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab73406, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
ab73406 has not yet been referenced specifically in any publications.
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