Anti-TREX1 antibody [EPR14985]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREX1 antibody [EPR14985] (AB185228)

Immunohistochemical analysis of paraffin embedded human colon adenocarcinoma tissue sections labeling TREX1 using ab185228 at a 1/100 dilution. Hematoxylin counterstain.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TREX1 antibody [EPR14985] (AB185228)

Confocal image showing cytoplasmic and weak nuclear staining in HeLa (human cervix adenocarcinoma epithelial cell) cells. Cells were fixed in 4% paraformaldehyde and permeabilised with 0.1% TritonX-100. The primary anti-TREX1 antibody, ab185228, was used at a 1 : 200 dilution (10 μg/ml). An AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) was used at a 1 : 1000 dilution (2 μg/ml) (green). An anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a counterstain at a 1 : 200 dilution (2.5 μg/ml) (red). DAPI (blue) was used as a nuclear counterstain. A secondary antibody only control was also performed.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TREX1 antibody [EPR14985] (AB185228)

Immunohistochemical analysis of paraffin embedded Human colon tissue sections labeling TREX1 using ab185228 at a 1/100 dilution. Hematoxylin counterstain.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-TREX1 antibody [EPR14985] (AB185228)

Lanes 1-4 : Merged signal (red and green). Green - ab185228 observed at 34 kDa. Red - loading control ab7291 observed at 50 kDa.

ab185228 Anti-TREX1 antibody [EPR14985] was shown to specifically react with TREX1 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266927 (knockout cell lysate ab257764) was used. Wild-type and TREX1 knockout samples were subjected to SDS-PAGE. ab185228 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TREX1 antibody [EPR14985] (ab185228) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TREX1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trex1-knockout-a549-cell-line-ab266927'>ab266927</a>)

Lane 2:

TREX1 knockout A549 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 34 kDa

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• WB

Supplier Data

Western blot - Anti-TREX1 antibody [EPR14985] (AB185228)

False colour image of Western blot : Anti-TREX1 antibody [EPR14985] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab185228 was shown to bind specifically to TREX1. A band was observed at 33 kDa in wild-type A549 cell lysates with no signal observed at this size in TREX1 knockout cell line ab266926 (knockout cell lysate ab257763). To generate this image, wild-type and TREX1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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.

All lanes:

Western blot - Anti-TREX1 antibody [EPR14985] (ab185228) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

TREX1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TREX1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-trex1-knockout-a549-cell-line-ab266926'>ab266926</a>)

Lane 3:

HEK-293T cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 39 kDa

Observed band size: 33 kDa

false

• WB

Supplier Data

Western blot - Anti-TREX1 antibody [EPR14985] (AB185228)

All lanes:

Western blot - Anti-TREX1 antibody [EPR14985] (ab185228) at 1/10000 dilution

Lane 1:

Raji cell lysate at 20 µg

Lane 2:

Daudi cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 39 kDa

false