Anti-IL-2 antibody [EPR16615-341]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-2 antibody [EPR16615-341] (AB243650)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized EL4.IL-2 (mouse lymphoma T lymphocyte) treated with cell stimulation cocktail (80nM PMA + 1.34uM Ionomycin + 10.6uM Brefeldin A + 2uM Monensin) for 6 hours (Red)/ Untreated control (Green) cells labelling IL-2 with ab243650 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IL-2 antibody [EPR16615-341] (AB243650)

Intracellular flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse splenocytes treated with cell stimulation cocktail (80nM PMA+1.34uM Ionomycin+10.6uM Brefeldin A+2uM Monensin) for 6 hours (Right)/ Untreated control (Left). cells labelling IL-2 with ab243650 at 1/500 dilution (1ug)/ Left and Right (Red) compared with a isotype control. A Goat anti rabbit IgG (Alexa Fluor^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody. Cells were surface stained with anti-CD3 conjugated to FITC. Then fixed with 2% PFA for 10min followed by intracellularly stained with ab243650. Gated on lymphocytes population. Data was kindly provided by an anonymous collaborator.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IL-2 antibody [EPR16615-341] (AB243650)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized EL4.IL-2 cells labelling IL-2 with ab243650 at 1/50 (9.1 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing cytoplasmic staining in EL4.IL-2 cell line treated with Cell Stimulation Cocktail (80nM PMA + 1.34uM Ionomycin + 10.6uM Brefeldin A + 2uM Monensin) for 6 h. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IL-2 antibody [EPR16615-341] (AB243650)

Flow cytometry staining of C57 BL/6 mouse splenocytes (top) or C57 BL/6 mouse splenocytes treated with 80nM PMA/TPA, 1.34uM Ionomycin and 1μg/mL Brefeldin A for 18 Hours (bottom) with ab243650 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Cells were incubated for 30min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab243650 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶in 100 μl at 1.0 μg/ml (1/500)) for 30min at 22°C. The cells were simultaneously stained with CD4.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.