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Recombinant full length Human SAMHD1 protein (NP_056289) produced in HEK293T cell.
Our Abpromise guarantee covers the use of ab117908 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.|
|WB||1/2000. Predicted molecular weight: 72 kDa.|
|Flow Cyt||1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: SAMHD knockout HAP1 cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lanes 1 - 3: Merged signal (red and green). Green - ab117908 observed at 70 kDa. Red - loading control, ab181602, observed at 37 kDa. ab117908 was shown to recognize SAMHD1 when SAMHD1 knockout samples were used, along with additional cross-reactive bands. Wild-type and SAMHD1 knockout samples were subjected to SDS-PAGE. ab117908 and ab181602 (loading control to GAPDH) were diluted 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
IHC image of SAMHD1 staining in Human normal spleen 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 ab117908, 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.
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.