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BALB/c macrophages obtained from 14-day-old bone marrow cell cultures.
ab16911 is the only macrophage marker that is able to distinguish non-destructive from destructive inflammation prcoesses in the pancreas. Furthermore it is a unique histological marker of the progression from peri-insulitis to beta-cell and diabetes in a mouse diabetes model.
Our Abpromise guarantee covers the use of ab16911 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
(Methanol fixed cells)
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|WB||1/50. Use under non reducing condition. Predicted molecular weight: 125 kDa.|
See Schaller et al.
Fixation with acetone for 10 min at RT is recommended as is an incubation with 0.02 M sodium azide in PBS containing 0.1 % H2O2 for 10 min at RT to destroy endogenous peroxidase
Paraffin embedded sections of mouse colon stained with ab16911 at 2 μg/ml.
Detection of F4/80 in RAW cells. Red, black and blue line represent the isotype control, cells only and ab16911 at 10 μg/ml, respectively.
ab16911 stained RAW246.7 cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16911 at 1in50 dilution) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (ab150165) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.