Anti-IL-2 antibody [EPR20055] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IL-2 antibody [EPR20055] - BSA and Azide free (AB251472)

This data was developed using ab207325, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells, untreated or treated with 40 nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight, labeling IL2 with ab207325 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on Jurkat cells. The signal is induced after treatment with 40 nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight.

The nuclear counter stain is DAPI (blue).

Tubulin is detected with (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only controls on non treated and treated cells : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-IL-2 antibody [EPR20055] - BSA and Azide free (AB251472)

This data was developed using ab207325, the same antibody clone in a different buffer formulation.

IL2 was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 40nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight lysate with ab207325 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab207325 at 1/200 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Jurkat treated with 40nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight lysate 10μg (Input).
Lane 2 : ab207325 IP in Jurkat treated with 40nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight lysate.
Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207325 in Jurkat treated with 40nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight lysate.

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-IL-2 antibody [EPR20055] (<a href='/en-us/products/primary-antibodies/il-2-antibody-epr20055-ab207325'>ab207325</a>)

Predicted band size: 17 kDa

false

• WB

Supplier Data

Western blot - Anti-IL-2 antibody [EPR20055] - BSA and Azide free (AB251472)

This data was developed using ab207325, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Human IL2 active recombinant protein contains aa21-153.

All lanes:

Western blot - Anti-IL-2 antibody [EPR20055] (<a href='/en-us/products/primary-antibodies/il-2-antibody-epr20055-ab207325'>ab207325</a>) at 1/1000 dilution

All lanes:

Human IL2 active recombinant protein at 0.01 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

Predicted band size: 17 kDa

Observed band size: 14 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-IL-2 antibody [EPR20055] - BSA and Azide free (AB251472)

This data was developed using ab207325, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the literature, and the expression level can be induced in activated Jurkat cell (PMID : 10555752).

All lanes:

Western blot - Anti-IL-2 antibody [EPR20055] (<a href='/en-us/products/primary-antibodies/il-2-antibody-epr20055-ab207325'>ab207325</a>) at 1/5000 dilution

Lane 1:

Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 2:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate treated with 40nM Phorbol-12-myristate-13-acetate, 2 μM A23187, and 300 ng/ml Brefeldin A (45 min delay) overnight at 10 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

Predicted band size: 17 kDa

Observed band size: 14 kDa

false

Exposure time: 1min