Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling SAMHD1 with ab177462 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

Intracellular flow cytometric analysis of permeabilized THP1 cells labeling SAMHD1 (red)with ab177462 at 1/10 dilution or rabbit IgG (negative) (green).

• WB

Supplier Data

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

All lanes:

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462) at 1/10000 dilution

Lane 1:

THP1 cell lysate at 10 µg

Lane 2:

K562 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

MCF7 cell lysate at 10 µg

Secondary

All lanes:

Goat-anti-rabbit HRP at 1/2000 dilution

Predicted band size: 72 kDa

true

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

Immunofluorescence analysis of MCF7 cells labeling SAMHD1 with ab177462 at 1/100 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling SAMHD1 with ab177462 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

Anti-SAMHD1 antibody [EPR13040(B)] (ab177462) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab177462 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

Lanes 1 - 5:

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462) at 1/10000 dilution

Lanes 1 - 5:

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/samhd1-antibody-epr13040b-bsa-and-azide-free-ab240199'>ab240199</a>)

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

Observed band size: 73 kDa

false

• WB

Lab

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (AB177462)

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 - ab177462 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab177462 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. ab177462 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SAMHD1 antibody [EPR13040(B)] - C-terminal (ab177462)

Predicted band size: 72 kDa

false