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Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Rat Hsp47.
Our Abpromise guarantee covers the use of ab86750 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 5 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 47 kDa).|
|Flow Cyt||Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Hsp47 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab86750 observed at 46 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab86750 was shown to recognize Hsp47 in wild-type HAP1 cells as signal was lost at the expected MW in Hsp47 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Hsp47 knockout samples were subjected to SDS-PAGE. ab86750 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 5 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
ab86750 has not yet been referenced specifically in any publications.