Anti-Transportin 1/MIP antibody [EPR29606-11]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Transportin 1/MIP with ab324273 at 1/50 (10.33 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing nuclear and cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324273 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

Transportin 1/MIP was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab324273 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324273 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Lane 2:

ab324273 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab324273 in HeLa whole cell lysate

Secondary

All lanes:

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

Observed band size: 102 kDa

false

Exposure time: 3s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Transportin 1/MIP with ab324273 at 1/50 (10.33 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing nuclear and cytoplasmic staining in NIH/3T3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324273 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution. -ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

Transportin 1/MIP was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab324273 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324273 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg

Lane 2:

ab324273 at 1/30 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab324273 in NIH/3T3 whole cell lysate

Secondary

All lanes:

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

Observed band size: 102 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

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

All lanes:

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 50 µg

Lane 2:

Human lymph node tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 102 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

HeLa transfected with siRNA specifically targeting Transportin 1/MIP whole cell lysate at 50 µg

Lane 3:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 50 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: 102 kDa,36 kDa

false

Exposure time: 59s

• Dot

Supplier Data

Dot Blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

Dot blot analysis of Transportin 1/MIP using ab324273 at 1 : 1000 (0.517 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : His-tagged humanTNPO2 recombinant protein
Lane2 : His-tagged humanTNPO1 recombinant protein

Exposure time : 180 seconds.

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

This antibody does not cross-react with human TNPO2.

In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Dot Blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

His-tagged humanTNPO2 recombinant protein

Lane 2:

His-tagged humanTNPO1 recombinant protein

Secondary

All lanes:

Dot 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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (AB324273)

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

Exposure time : Lanes 1-2 : 103 seconds, Lanes 3-4 : 37 seconds

All lanes:

Western blot - Anti-Transportin 1/MIP antibody [EPR29606-11] (ab324273) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 50 µg

Lane 2:

Mouse cerebellum tissue lysate at 50 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 50 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 50 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

Observed band size: 102 kDa

false