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Our Abpromise guarantee covers the use of ab25625 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 0.01µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|Neutralising||Use at an assay dependent concentration. PubMed: 21789185|
Flow Cytometry analysis of Human monocyte cells labeling CD14 with ab25625 at 1 μg/106 cells (purple). A Goat Anti-Mouse IgG1, Human ads-CY5 was used as the secondary antibody. Grey - Isotype Control, Mouse IgG1-UNLB, followed by Goat Anti-Mouse IgG1, Human ads-CY5.
Human peripheral blood cells stained with ab25625 (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 (ab25625, 0.01μg/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. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Cells were incubated for 1 hr with 10 µg/ml ab25625 prior to addition of 100 ng/ml LPS for 24 hrs. The corresponding IgG isotypes were used as negative controls.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"