Anti-Emerin antibody [EPR11071]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Emerin antibody [EPR11071] (AB156871)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling Emerin with unpurified ab156871 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Emerin antibody [EPR11071] (AB156871)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue sections labeling Emerin with purified ab156871 at 1/500 dilution (0.652 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Emerin antibody [EPR11071] (AB156871)

Immunohistochemical analysis of paraffin-embedded Human thyroid gland carcinoma tissue labeling Emerin with unpurified ab156871 at 1/50 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Emerin antibody [EPR11071] (AB156871)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells, labeling Emerin with Purified ab156871 at 1/30 dilution (10μg/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). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Emerin antibody [EPR11071] (AB156871)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Emerin with Purified ab156871 at 1 : 50 dilution (6.5 μg/ml). Cells were fixed in 4% Paraformaldehyde 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.

• WB

Lab

Western blot - Anti-Emerin antibody [EPR11071] (AB156871)

Lanes 1 - 3 : Merged signal (red and green). Green - ab156871 observed at 32 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab156871 was shown to specifically react with Emerin in wild-type HAP1 cells as signal was lost in EMD (Emerin) knockout cells. Wild-type and EMD (Emerin) knockout samples were subjected to SDS-PAGE. ab156871 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 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-Emerin antibody [EPR11071] (ab156871) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

EMD (Emerin) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 29 kDa

false

• WB

Lab

Western blot - Anti-Emerin antibody [EPR11071] (AB156871)

Lanes 1-2 : Merged signal (red and green). Green - ab156871 observed at 35 kDa. Red - loading control ab8245 observed at 37 kDa.

ab156871 Anti-Emerin antibody [EPR11071] was shown to specifically react with Emerin in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266336 (knockout cell lysate ab257423) was used. Wild-type and Emerin knockout samples were subjected to SDS-PAGE. ab156871 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-Emerin antibody [EPR11071] (ab156871) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

EMD knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human EMD (Emerin) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-emd-emerin-knockout-hek-293t-cell-line-ab266336'>ab266336</a>)

Predicted band size: 29 kDa,51 kDa,72 kDa

Observed band size: 35 kDa,51 kDa

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• WB

Unknown

Western blot - Anti-Emerin antibody [EPR11071] (AB156871)

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Emerin antibody [EPR11071] (ab156871) at 1/10000 dilution

Lane 1:

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

Lane 2:

HEK-293 (Human embryonic kidney epithelial cell) 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 1/20000 dilution

Predicted band size: 29 kDa

Observed band size: 35 kDa

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