Anti-IL-2 Receptor alpha antibody [OX39] - BSA and Azide free

2 product images

• 

  Flow Cytometry - Anti-IL-2 Receptor alpha antibody [OX39] - BSA and Azide free (ab244557)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-IL-2 Receptor alpha antibody [OX39] ab6411).
  

  Lewis rat splenocytes (top) or Lewis rat splenocytes treated with 5 μg/ml ConA for 3 days (bottom) were stained with Anti-IL-2 Receptor alpha antibody [OX39] ab6411 (right) or mouse IgG1 kappa (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) isotype (left).

  Splenocytes were incubated for 30 min on ice in 1x PBS containing 10 % rat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (Anti-IL-2 Receptor alpha antibody [OX39] ab6411) or mouse IgG1 kappa (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) isotype (1x 10⁶ in 100 μl at 0.2 μg/ml) for 30 min on ice.

  The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 dilution for 30 min on ice.

  The cells were simultaneously stained with CD4.

  Acquisition of >30,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on CD3 positive T cells.

• 

  Immunohistochemistry (Frozen sections) - Anti-IL-2 Receptor alpha antibody [OX39] - BSA and Azide free (ab244557)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-IL-2 Receptor alpha antibody [OX39] ab6411).
  

  IHC image of IL2 receptor alpha staining in a section of frozen normal rat spleen.

  The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with Anti-IL-2 Receptor alpha antibody [OX39] ab6411 at 5μg/ml and Anti-CD3 epsilon antibody [SP7] ab16669 (Rabbit monoclonal [SP7] to CD3) at 1/150. The section was then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor^®488), 1/1000)) (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594), 1/1000) (shown in red) for 1 hour at room temperature. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM. The secondary-only control insert image is taken from an identical assay without primary antibody. The section was then mounted using Fluoromount^®.

  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.