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Our Abpromise guarantee covers the use of ab56297 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 2µg for 106 cells.
ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
|IHC-P||Use at an assay dependent concentration. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.|
ICC/IF image of ab56297 stained RAW246.7 cells. The cells were 100% methanol fixed (5 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 (ab56297, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96887, 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
Ab56297 staining macrophage in mouse brain tumour tissue sections by (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 0.5% TNB blocking reagent for 30 minutes at 25°C. Samples were incubated with primary antibody at 1/100 dilution for 18 hours at 4 C. A goat anti-rat IgG H&L (HRP) (ab7097) was used at 1/500 dilution.
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