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Our Abpromise guarantee covers the use of ab6173 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-FoFr||1/500. PubMed: 19200078|
ab6173 staining Neuropeptide Y in rat brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/10000) for 16 hours at 4°C. A Biotin-conjugated Donkey anti-sheep IgG polyclonal (1/500) was used as the secondary antibody.
ICC/IF image of ab6173 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal donkey 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 ab6173 at 1/100 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 donkey anti- sheep (ab96939) 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.