Anti-Nesprin 2 antibody [EPR28137-54]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Nesprin 2 with ab314872 at 1/200 (2.58 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on nuclear envelope in human tonsil. The section was incubated with ab314872 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Immunohistochemical analysis of paraffin-embedded Human gastric adenocarcinoma tissue labeling Nesprin 2 with ab314872 at 1/200 (2.58 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on nuclear envelope in human gastric adenocarcinoma. The section was incubated with ab314872 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Immunohistochemical analysis of paraffin-embedded Human endometrial carcinoma tissue labeling Nesprin 2 with ab314872 at 1/200 (2.58 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on nuclear envelope in human endometrial carcinoma. The section was incubated with ab314872 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Nesprin 2 with ab314872 at 1/50 (10.32 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear envelope staining in 293T cells and weak nuclear staining in THP-1 cells. Low expression : THP-1. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• WB

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Western blot - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Nesprin 2 has several splicing variants. The molecular weights are consistent with what have been described in literatures(PMID : 15843432, PMID : 24718612, PMID : 18477613). Exposure time : Lane1 : 37 seconds, Lane2 : 125 seconds

All lanes:

Western blot - Anti-Nesprin 2 antibody [EPR28137-54] (ab314872) at 1/1000 dilution

Lane 1:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HaCaT (human skin keratinocyte) 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

Observed band size: 280 kDa

false

• WB

Supplier Data

Western blot - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Nesprin 2 has several splicing variants. The molecular weights are consistent with what have been described in literatures(PMID : 15843432, PMID : 24718612, PMID : 18477613). Low expression : THP-1 In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Nesprin 2 antibody [EPR28137-54] (ab314872) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

THP-1 (human monocytic leukemia monocyte) 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

Observed band size: 280 kDa

false

Exposure time: 59s

• WB

Lab

Western blot - Anti-Nesprin 2 antibody [EPR28137-54] (AB314872)

Western blot : Anti-SYNE2 antibody [EPR28137-54] (ab314872) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab314872 was shown to bind specifically to SYNE2. A band was observed at 796 kDa in wild-type A549 cell lysates with no signal observed at this size in SYNE2 knockout cell line. To generate this image, wild-type and SYNE2 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 3 % 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-Nesprin 2 antibody [EPR28137-54] (ab314872) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SYNE2 knockout A549 cell lysate at 20 µg

Lane 3:

U-2 OS cell lysate at 20 µg

Lane 4:

THP-1 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

Observed band size: 796 kDa

false