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Tissue, cells or virus corresponding to Mouse CD68.
Although some customers have had success with this antibody in IHC-P, we are unable to obtain positive results in this application and so cannot recommend it for IHC-P. We batch test the antibody in IHC-Fr.
This antibody clone is manufactured by Abcam.
Our Abpromise guarantee covers the use of ab53444 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 5 µg/ml.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use a concentration of 0.1 - 5 µg/ml.|
|Flow Cyt||1/50 - 1/100.
Use 10µl of the suggested working dilution to label 106 cells in 100µl. Membrane permeabilisation is required for this application.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ab53444 stained RAW246.7 cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature 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 (ab53444 at 5µg/ml) 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.
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.
ab53444 staining mouse heart tissue by Immunohistochemistry (IHC-frozen sections). Tissue underwent fixation in paraformaldehyde, no permeabilization and blocked in 1% serum for 10 minutes at 20°C. The primary antibody used undiluted and incubated with sample for 16 hours at 20°C. A Biotin conjugated goat polyclonal to rat Ig, diluted 1/100 was used as the secondary.
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