Anti-DEK antibody [EPR20401]

• WB

Supplier Data

Western blot - Anti-DEK antibody [EPR20401] (AB221545)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-2 : 2 seconds; Lane 3 : 3 seconds.

Lanes 1 - 2:

Western blot - Anti-DEK antibody [EPR20401] (ab221545) at 1/2000 dilution

Lane 3:

Western blot - Anti-DEK antibody [EPR20401] (ab221545) at 1/10000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 2:

293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 3:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell 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 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 50 kDa

true

• WB

Lab

Western blot - Anti-DEK antibody [EPR20401] (AB221545)

Western blot : Rabbit Monoclonal[EPR20401] to DEK ab221545 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 52 kDa in Wild-type HeLa ab271142 cell lysates with no signal observed at this size in DEK knockout HeLa ab274942 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 [EPR20401] (ab221545) at 1/2000 dilution

Lane 1:

Wild-type HeLa ab271142 at 20 µg

Lane 2:

Western blot - Human DEK knockout HeLa cell line (ab274942) 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: 52 kDa

Observed band size: 52 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DEK antibody [EPR20401] (AB221545)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR20401] (AB221545)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human colon is observed (PMID : 23902796). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-DEK antibody [EPR20401] (AB221545)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DEK antibody [EPR20401] (ab221545) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 10 µg

Lane 2:

Human fetal heart lysate at 10 µg

Lane 3:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/4000 dilution

Predicted band size: 42 kDa

Observed band size: 50 kDa

true

Exposure time: 3min

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR20401] (AB221545)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong nuclear staining on human colon cancer is observed (PMID : 23902796). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR20401] (AB221545)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human gastric cancer is observed (PMID : 26998084). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR20401] (AB221545)

Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Nuclear staining on human lung cancer is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DEK antibody [EPR20401] (AB221545)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Weak cytoplasmic and strong nuclear staining on human breast cancer is observed (PMID : 26459608). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-DEK antibody [EPR20401] (AB221545)

DEK was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab221545 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221545 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).

Lane 2 : ab221545 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221545 in HeLa whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-DEK antibody [EPR20401] (ab221545)

Predicted band size: 42 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DEK antibody [EPR20401] (AB221545)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HEK-293T cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DEK antibody [EPR20401] (AB221545)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized K562 (human chronic myelogenous leukemia cell line from bone marrow) cells labeling DEK with ab221545 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on K562 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-DEK antibody [EPR20401] (AB221545)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed K562 (human chronic myelogenous leukemia cell line from bone marrow) cell line labeling DEK with ab221545 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.