Anti-DDX17 antibody [EPR13807(B)]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling DDX17 with Purified ab180190 at 1 : 50 dilution (2 μg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Flow Cytometry analysis of J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage, Left) / NIH/3T3 (Mouse embryonic fibroblast, Right) cells labelling DDX17 with Purified ab180190 at 1 : 20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling DDX17 with Purified ab180190 at 1 : 5000 dilution (0.02 μg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

Lab

Western blot - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Lanes 1 - 4 : Merged signal (red and green). Green - ab180190 observed at 72 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab180190 was shown to recognize DDX17 in wild-type HEK 293 cells as signal was lost at the expected MW in DDX17 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and DDX17 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab180190 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-DDX17 antibody [EPR13807(B)] (ab180190) at 1/5000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

DDX17 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human DDX17 knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-ddx17-knockout-hek-293-cell-line-ab261721'>ab261721</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 72 kDa

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• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling DDX17 with Purified ab180190 at 1 : 5000 dilution (0.02 μg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling DDX17 with Purified ab180190 at 1 : 5000 dilution (0.02 μg/ml). Heat mediated antigen retrieval was performed using . Tissue was counterstained with Hematoxylin. Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0) secondary antibody was used at 1 : 0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

Unknown

Western blot - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

All lanes:

Western blot - Anti-DDX17 antibody [EPR13807(B)] (ab180190) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Rat brain lysate at 20 µg

Lane 4:

Rat testis lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 72 kDa

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• WB

Lab

Western blot - Anti-DDX17 antibody [EPR13807(B)] (AB180190)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 6 s

All lanes:

Western blot - Anti-DDX17 antibody [EPR13807(B)] (ab180190) at 1/1000 dilution

Lane 1:

HeLa (Human colorectal adenocarcinoma epithelial cell) cell lysate at 10 µg

Lane 2:

Mouse embryo lysate at 10 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) cell lysate at 10 µg

Lane 4:

J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) cell lysate at 10 µg

Lane 5:

RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cell lysate at 10 µg

Lane 6:

C2C12 (Mouse myoblasts myoblast) cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 72 kDa

Observed band size: 72 kDa

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