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Synthetic peptide conjugated to KLH derived from within residues 600 - 700 of Human Integrin alpha 5.
Our Abpromise guarantee covers the use of ab112183 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 1 µg/ml. Detects a band of approximately 145 kDa (predicted molecular weight: 114 kDa).|
|IHC-P||Use a concentration of 1 µg/ml.|
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Integrin alpha 5 knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jeg3 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab112183 observed at 140 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab112183 was shown to recognize Integrin alpha 5 when Integrin alpha 5 knockout samples were used, along with additional cross-reactive bands. Wild-type and Integrin alpha 5 knockout samples were subjected to SDS-PAGE. Ab112183 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
IHC image of Integrin alpha 5 staining in Human normal placenta 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 ab112183, 1µg/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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab112183 has not yet been referenced specifically in any publications.