Anti-SAMHD1 antibody [OTI1F9]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

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).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

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.

• WB

Lab

Western blot - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

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.

All lanes:

Western blot - Anti-SAMHD1 antibody [OTI1F9] (ab117908)

Predicted band size: 72 kDa

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• WB

Supplier Data

Western blot - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

All lanes:

Western blot - Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/200 dilution

Lane 1:

HepG2 (human liver hepatocellular carcinoma cell line) cell extract at 10 µg

Lane 2:

HeLa (human epithelial cell line from cervix adenocarcinoma) cell extract at 10 µg

Lane 3:

SVT2 cell extract at 10 µg

Lane 4:

Jurkat (human T cell leukemia cell line from peripheral blood) cell extract at 10 µg

Lane 5:

MCF7 (human breast adenocarcinoma cell line) cell extract at 10 µg

Predicted band size: 72 kDa

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• WB

Lab

Western blot - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

Western blot : Anti-SAMHD1 antibody [OTI1F9] (ab117908) staining at 1/2000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab117908 was shown to bind specifically to SAMHD1. A band was observed at 73 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in SAMHD1 knockout cell line. To generate this image, wild-type and SAMHD1 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/2000 dilution

Lane 1:

HEK-293 cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 3:

Wild-type HAP1 cell lysate at 20 µg

Lane 4:

SAMHD1 knockout HAP1 cell lysate at 20 µg

Lane 5:

Human Spleen cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Predicted band size: 72 kDa

Observed band size: 73 kDa

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• WB

Supplier Data

Western blot - Anti-SAMHD1 antibody [OTI1F9] (AB117908)

All lanes:

Western blot - Anti-SAMHD1 antibody [OTI1F9] (ab117908) at 1/200 dilution

All lanes:

Mouse brain extract at 10 µg

Predicted band size: 72 kDa

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