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The details of the immunogen for this antibody are not available.
Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab210219 as a replacement.
Our Abpromise guarantee covers the use of ab25235 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 a concentration of 10 µg/ml.|
|Flow Cyt||Use 3µg for 106 cells.
ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|Blocking||Use 2µg for 106 cells.
ab25235 can be used for blocking of Fc gamma receptors.
ICC/IF image of ab25235 stained RAW246.7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab25235, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab98420, DyLight® 488 goat anti-rat IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Flow Cytometry analysis of BALB/c mouse splenocytes cells labeling CD16/32 with ab25235 at 1 μg/106 cells dilution. A Rat Anti-Mouse CD19-FITC was used as the secondary antibody.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"