Anti-Importin subunit alpha antibody [EPR26346-53]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Importin subunit alpha with ab307438 at 1/50 dilution (10.02 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic and nuclear staining in untreated HeLa cells, and the signals were translocated to nucleus in HeLa cells after treatment with heat shock (45.5°C) for 1.5 h. 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 dilution (2.5 ug/ml) (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 dilution (2 ug/ml).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Importin subunit alpha with ab307438 at 1/2000 dilution (0.251 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human breast (PMID : 34709479). The section was incubated with ab307438 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is 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 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Immunohistochemical analysis of paraffin-embedded human colorectal adenocarcinoma tissue labeling Importin subunit alpha with ab307438 at 1/2000 dilution (0.251 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human colorectal adenocarcinoma. The section was incubated with ab307438 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is 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 20 mins.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Importin subunit alpha with ab307438 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling Importin subunit alpha with ab307438 at 1/2000 dilution (0.251 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining on human breast cancer (PMID : 34709479). The section was incubated with ab307438 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is 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 20 mins.

• IP

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Immunoprecipitation - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Importin subunit alpha was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab307438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307438 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HepG2 whole cell lysate Lane 2 : ab307438 IP in HepG2 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307438 in HepG2 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-Importin subunit alpha antibody [EPR26346-53] (ab307438) at 1/1000 dilution

All lanes:

HepG2 whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Importin subunit alpha with ab307438 at 1/50 dilution (10.02 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic and nuclear staining in untreated NIH/3T3 cells, and the signals were translocated to nucleus in NIH/3T3 cells after treatment with heat shock (45.5°C) for 1.5 h. 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 dilution (2.5 ug/ml) (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 dilution (2 ug/ml).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Importin subunit alpha with ab307438 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 39 seconds.

All lanes:

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (ab307438) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting SRP1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Exposure time: 39s

• WB

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Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (ab307438) at 1/1000 dilution

Lane 1:

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

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 147 seconds.

All lanes:

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (ab307438) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Exposure time: 147s

• WB

Supplier Data

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (AB307438)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 37 seconds.

All lanes:

Western blot - Anti-Importin subunit alpha antibody [EPR26346-53] (ab307438) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Rat brain tissue lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Exposure time: 37s