Anti-DEK antibody [EPR11034]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR11034] (AB166624)

Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling DEK with ab166624 at 1/250 dilution.

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-DEK antibody [EPR11034] (AB166624)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling DEK (red) with ab166624 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-DEK antibody [EPR11034] (AB166624)

Immunofluorescence analysis of 293T cells labeling DEK with ab166624 at 1/100 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR11034] (AB166624)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling DEK with ab166624 at 1/250 dilution.

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

• IP

Unknown

Immunoprecipitation - Anti-DEK antibody [EPR11034] (AB166624)

Immunoprecipitation of DEK in pellet from HeLa cell lysate using ab166624 at 1/10 dilution, followed by WB detection of DEK using as secondary an HRP-conjugated anti-rabbit IgG, which preferentially detects the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-DEK antibody [EPR11034] (ab166624) at 1/1000 dilution

All lanes:

immunoprecipitation pellet from HeLa cell lysate at 10 µg

Predicted band size: 42 kDa

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• WB

Lab

Western blot - Anti-DEK antibody [EPR11034] (AB166624)

Western blot : Rabbit Monoclonal[EPR11034] to DEK ab166624 staining at 1/2000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 50 kDa in Wild-type HeLa cell lysates with no signal observed at this size in DEK knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-DEK antibody [EPR11034] (ab166624) at 1/2000 dilution

Lane 1:

Wild-type HeLa at 20 µg

Lane 2:

Human DEK knockout HeLa cell line at 20 µg

Lane 3:

T-47D at 20 µg

Lane 4:

K562 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

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• WB

Unknown

Western blot - Anti-DEK antibody [EPR11034] (AB166624)

All lanes:

Western blot - Anti-DEK antibody [EPR11034] (ab166624) at 1/1000 dilution

Lane 1:

293T cell lysates at 10 µg

Lane 2:

HeLa cell lysates at 10 µg

Lane 3:

Jurkat cell lysates at 10 µg

Lane 4:

K562 cell lysates at 10 µg

Predicted band size: 42 kDa

false