Anti-FOXP2 antibody [EPR29629-38]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized SK-N-MC (human brain epithelial cell) cells labelling FOXP2 with ab326725 at 1/200 (2.495 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in SK-N-MC cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled 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 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326725 at 1/200 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MEF (mouse embryo fibroblast) cells labelling FOXP2 with ab326725 at 1/200 (2.495 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in MEF cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled 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 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326725 at 1/200 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

FOXP2 was immunoprecipitated from 0.35 mg MEF (mouse embryo fibroblast) whole cell lysate with ab326725 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326725 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MEF (mouse embryo fibroblast) whole cell lysate
Lane 2 : ab326725 IP in MEF (mouse embryo fibroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326725 in MEF whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-FOXP2 antibody [EPR29629-38] (ab326725) at 1/1000 dilution

Lane 1:

MEF (mouse embryo fibroblast) whole cell lysate at 10 µg

Lane 2:

ab326725 at 1/30 IP in MEF (mouse embryo 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 ab326725 in MEF 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: 80 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

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

Exposure time : Lane1 : 59 seconds, lane2 : 180 seconds

All lanes:

Western blot - Anti-FOXP2 antibody [EPR29629-38] (ab326725) at 1/1000 dilution

Lane 1:

SK-N-MC (human brain epithelial cell) fresh whole cell lysate at 20 µg

Lane 2:

MEF (mouse embryo fibroblast) fresh 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

Observed band size: 80 kDa

false

• WB

Lab

Western blot - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-FOXP2 antibody [EPR29629-38] (ab326725) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control fresh whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting FOXP2 fresh 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

Observed band size: 80 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-FOXP2 antibody [EPR29629-38] (AB326725)

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

Low expression : testis

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) 1 : 10000 (124Kda).

All lanes:

Western blot - Anti-FOXP2 antibody [EPR29629-38] (ab326725) at 1/1000 dilution

Lane 1:

Mouse E12 embryo fresh tissue lysate at 60 µg

Lane 2:

Mouse E15 embryo fresh tissue lysate at 60 µg

Lane 3:

Mouse testis fresh tissue lysate at 60 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: 80 kDa,124 kDa

false

Exposure time: 92s