Staining of a cytospin preparation of peripheral blood mononuclear cells isolated from buffycoat. Cells were left to air dry and then fixed with cold acetone (90 seconds) and blocked with PBS containing 1% FCS and 0.1% saponin (blocking buffer) for 20 minutes. Cells were then washed twice in PBS and incubated with Sheep antibody to human Cathelicidin antimicrobial peptide: whole serum (OSC00037W), diluted 1 : 200 in blocking buffer, for 1 hour at room temperature. Cells were washed twice in PBS followed by incubation for 30 minutes with the FITC-labeled secondary antibody. Finally, cells were washed and nuclear counterstained with DAPI.  The antibody selectively recognizes polymorphonuclar cells.

Staining of a cytospin preparation of peripheral blood mononuclear cells isolated from buffycoat. Cells were left to air dry and then fixed with cold acetone (90 seconds) and blocked with PBS containing 1% FCS and 0.1% saponin (blocking buffer) for 20 minutes. Cells were then washed twice in PBS and incubated with Sheep antibody to human Cathelicidin antimicrobial peptide: whole serum (OSC00037W), diluted 1 : 200 in blocking buffer, for 1 hour at room temperature. Cells were washed twice in PBS followed by incubation for 30 minutes with the FITC-labeled secondary antibody. Finally, cells were washed and nuclear counterstained with DAPI. The antibody selectively recognizes polymorphonuclar cells.

WB on human neutrophil lysate using Sheep antibody to human Cathelicidin antimicrobial peptide (CAP-18, hCAP-18, antibacterial protein LL-37, CAMP, CRAMP, FALL39): whole serum (OSC00037W) at a dilution of 1 : 100. Pre-absorption of the antibody with the immunising peptide completely abolishes the detected band.

IHC-P on paraffin sections of human large intestine tissue. HIER: Tris-EDTA, pH 9.0 for 20 min using Thermo PT Module. Blocking: 0.2% LFDM in TBST filtered thru 0.2 µm. Detection was done using Novolink HRP polymer from Leica following manufacturer's instructions. Primary antibody: dilution 1: 1000, incubated 30 min at RT (using Autostainer). Sections were counterstained with Harris Hematoxylin.