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This antibody was raised against a synthetic peptide derived from the N-terminus of C5a (amino acids 1-31).
Our Abpromise guarantee covers the use of ab11867 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Inhibition Assay||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 42 kDa).|
|Flow Cyt||Use 1µg for 106 cells. ab170191-Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.|
|Functional Studies||Use at an assay dependent concentration.|
|IHC-P||Use at an assay dependent concentration.|
Human peripheral blood granulocytes stained with ab11867 (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 (ab11867, 1μg/1x106 cells) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 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 granulocytes.