The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use 3µg for 106 cells. ab18536-Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
Use 2µg for 106 cells.
ab25235 can be used for blocking of Fc gamma receptors.
RelevanceFunction: Binds to the Fc region of immunoglobulins gamma. Low affinity receptor. By binding to IgG it initiates cellular responses against pathogens and soluble antigens. Promotes phagocytosis of opsonized antigens.
Tissue specificity: Found on monocytes, neutrophils and eosinophil platelets.
Similarity: Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
PTM: Phosphorylated by SRC-type Tyr-kinases such as LYN, BLK, FYN, HCK and SYK.
Cellular localizationCD32: Type I membrane protein.
CD16: Attached to the membrane by a GPI anchor. Exists also as a soluble receptor, produced by a proteolytic cleavage.
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.