ab53444 stained in RAW 246.7 cells. Cells were fixed with 100% methanol (5 min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% Tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab53444 at 1µg/ml and ab7291 (Mouse monoclonal to alpha Tubulin - Loading Control) used at a 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150165, Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed (pseudo-colored green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), preadsorbed (colored red), both used at 1/1000 dilution for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1hour at room temperature.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab53444).

Formaldehyde-fixed, frozen mouse lung tissue sections stained for CD68 using ab53444 at a 1/250 dilution in immunohistochemical analysis. Tissue sections were blocked using 1% BSA as a blocking agent for 10 minutes at 21°C. Primary antibody was incubated for 2 hours at 21°C. Secondary antibody was a biotin-conjugated goat anti-rat IgG at 1/250 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab53444).

IHC image of CD68 staining in mouse lung frozen tissue section. The section was incubated with ab53444, 0.1µg/ml, overnight at 4C. A goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab53444).

ab53444 staining CD68 in mouse spleen tissue by Immunohistochemistry (Frozen sections). Antibody was detected with HRP-conjugated Goat anti-Rat IgG, showing staining in the red pulp.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab53444).

Staining of permeabilised mouse peritoneal macrophages with Rat anti mouse CD68 (ab53444).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab53444).