Alexa Fluor® 488 Anti-CD8 alpha antibody [EPR21769]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Alexa Fluor® 488 Anti-CD8 alpha antibody [EPR21769] (AB237364)

IHC image of CD8 alpha staining in a section of formalin-fixed paraffin-embedded normal mouse spleen.

The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH9, epitope retrieval solution 2) for 20mins, performed on a Leica BOND^™. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) 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 ab237364 at 1/100 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®.

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

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

• Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Anti-CD8 alpha antibody [EPR21769] (AB237364)

C57BL/6 mouse splenocytes stained with ab237364 (right) or Rabbit IgG Alexa Fluor 488^® (monoclonal) (left). C57BL/6 Mouse splenocytes were incubated for 30 mins on ice in 1x PBS containing 10 μg/ml anti CD16/32 antibody and 10% normal goat serum to block FC receptors and non-specific protein-protein interactions followed by the antibody (ab237364) or Rabbit IgG 488 Isotype (1x10⁶ in 100 μl at 1 : 500 (1 μg/ml)) for 30 mins on ice. The cells were simultaneously stained with CD4 PerCP conjugated antibody.

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