Anti-IL23A antibody [EPR25909-4]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL23A antibody [EPR25909-4] (AB324195)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1(mouse reticulum cell sarcoma monocyte/macrophage) cells labelling IL23A with ab324195 at 1/1000 (0.514 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 increased cytoplasmic staining in J774A.1 cells treated with 100 ng/ml lipopolysaccharide (LPS) for 6 hours, with addition of brefeldin A (1 ug/ml) for the last 3 hours (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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta).

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL23A antibody [EPR25909-4] (AB324195)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling IL23A with ab324195 at 1/1000 (0.514 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 increased cytoplasmic staining in Raw264.7 cells treated with 100ng/ml lipopolysaccharide (LPS) for 6 hours, with addition of brefeldin A (1 ug/ml) for the last 3 hours (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).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta).

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.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL23A antibody [EPR25909-4] (AB324195)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 100ng/ml LPS for 6hours then 1ug/ml BFA for 3hours (Green) / Untreated control (Magenta) cells labelling IL23A with ab324195 at 1/500 dilution (0.1ug) (Magenta and Green) compared with a Rabbit monoclonal IgG (ab172730) (Black) and (Grey) isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-IL23A antibody [EPR25909-4] (AB324195)

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-IL23A antibody [EPR25909-4] (ab324195) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) transfected with scrambled siRNA control, then treated with 100ng/ml LPS(Lipopolysaccharide) for 3 hours, 300ng/ml Brefeldin A/BFA was then added for additional 3 hours whole cell lysate at 20 µg

Lane 2:

RAW 264.7 transfected with siRNA specifically targeting IL23A, then treated with 100ng/ml LPS(Lipopolysaccharide) for 3 hours, 300ng/ml Brefeldin A/BFA was then added for additional 3 hours 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: 36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL23A antibody [EPR25909-4] (AB324195)

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

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-IL23A antibody [EPR25909-4] (ab324195) at 1/1000 dilution

Lane 1:

Untreated J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate at 20 µg

Lane 2:

J774A.1 treated with 100ng/ml LPS(Lipopolysaccharide) for 3 hours, 300ng/ml Brefeldin A/BFA was then added for additional 3 hours 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: 36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL23A antibody [EPR25909-4] (AB324195)

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-IL23A antibody [EPR25909-4] (ab324195) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg

Lane 2:

RAW 264.7 treated with 100ng/ml LPS(Lipopolysaccharide) for 3 hours, 300ng/ml Brefeldin A/BFA was then added for additional 3 hours whole cell lysate at 40 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: 36 kDa

false

Exposure time: 92s