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 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed ab216777 secondary antibodies at 1/10000 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.

All lanes : Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/2000 dilution

Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate transfected with pCMV6-ENTRY control cDNA
Lane 2 : HEK-293T cell lysate transfected with pCMV6-ENTRY SAMHD1 cDNA

Lysates/proteins at 5 µg per lane.

Predicted band size: 72 kDa

All lanes : Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/200 dilution

Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) cell extract
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell extract
Lane 3 : SVT2 cell extract
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) cell extract
Lane 5 : MCF7 (human breast adenocarcinoma cell line) cell extract

Lysates/proteins at 10 µg per lane.

Predicted band size: 72 kDa

Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/200 dilution + Mouse brain extract at 10 µg

Predicted band size: 72 kDa

Flow cytometric analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for SAMHD1 using ab117908 (red) at 1/100 dilution, compared to a nonspecific negative control (blue).