Recombinant Anti-CD8 alpha antibody [SP16] (ab101500)

Immunohistochemical analysis of paraffin-embedded Human tonsil labeling CD8 with ab101500 at 1/100 (21 µg/ml). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab101500 for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. DAB was used as the chromogen. Counterstained with Hematoxylin and mounted with DPX.

This image was generated using a previous batch manufactured using hybridoma production method.

IHC using multi-spectral imaging on human lymph node (A-C) obtained from men with mCRPC. A) H+E staining and B) single-color images (plus nuclear stain; DAPI) of CD3 (ab16669), CD8 alpha (ab101500), CD163, PD-L1, cytokeratin (CK), DAPI and C) merged.  H+E stating at 20X magnification; multi-spectral images 200X magnification.

This image was generated using a previous batch manufactured using hybridoma production method.

Human peripheral blood lymphocytes staining CD8 alpha with ab101500 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab101500, 1/1000 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.