Anti-IL-2 antibody [RM1124]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-2 antibody [RM1124] (AB317331)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling IL-2 with ab317331 at 1/50 (10.0 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 cytoplasmic staining in jurkat cell line (shown in green) treated with 12-O-Tetradecanoylphorbol-13-acetate (15 nM), ionomycin (1 μm) and Brefeldin A (300 ng/mL) for 24 hr. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Left Panel is applied with Anti-IL-2 antibody [RM1024] (ab317331) at 1/50 dilution (10 ug/ml); Right panel is applied with Anti-IL-2 antibody (ab9618) at 1/100 dilution (10 ug/ml).
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 (Magenta). 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.

• IP

Supplier Data

Immunoprecipitation - Anti-IL-2 antibody [RM1124] (AB317331)

IL-2 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate with ab317331 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317331 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate
Lane 2 : ab317331 IP in Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317331 in Jurkat whole cell lysate

All lanes:

Immunoprecipitation - Anti-IL-2 antibody [RM1124] (ab317331) at 1/30 dilution

All lanes:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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

false

Exposure time: 180s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-2 antibody [RM1124] (AB317331)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling IL-2 with ab317331 at 1/50 (10.0 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 cytoplasmic staining in EL4 cells (shown in green) treated with TPA (40 nM), A23187 (2 uM) and BFA (300 ng/ml) for 23 hr. 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).

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

Supplier Data

Western blot - Anti-IL-2 antibody [RM1124] (AB317331)

The expression of IL-2 is upregulated in response to PMA/ionomycin treatment (PMID : 23494519).

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

All lanes:

Western blot - Anti-IL-2 antibody [RM1124] (ab317331) at 1/1000 dilution

Lane 1:

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

Lane 2:

Jurkat treated with 15nM 24h PMA (<a href='/en-us/products/biochemicals/phorbol-12-myristate-13-acetate-pma-pkc-activator-ab120297'>ab120297</a>) and 1µM ionomycin (<a href='/en-us/products/biochemicals/ionomycin-ca2-salt-ca2-ionophore-ab120116'>ab120116</a>) and 300ng/ml BFA for 24 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: 15 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-IL-2 antibody [RM1124] (AB317331)

The expression of IL-2 is upregulated in response to PMA/ionomycin treatment (PMID : 23494519).

The identity of the higher MW bands at approximately 50kDa are unknown.

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

All lanes:

Western blot - Anti-IL-2 antibody [RM1124] (ab317331) at 1/1000 dilution

Lane 1:

Untreated EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 2:

EL4.IL-2 cell line treated with Cell Stimation Cocktail (80nM PMA + 1.34uM Ionomycin + 10.6uM Brefeldin A + 2uM Monensin) for 6 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: 17 kDa,36 kDa

true

Exposure time: 103s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-IL-2 antibody [RM1124] (AB317331)

Indirect ELISA analysis of ab317331 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Human IL-2,Mouse IL-2.

Antigen concentration : 1000 ng/ml