ab14184 staining Substance P in mouse spinal cord tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

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ICC/IF image of ab14184 stained PC12 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 (ab14184, 1ug/ml) overnight at +4°C. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) used at a 1/1000 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 1.43uM.

ab14184 staining Substance P in human pancreas tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in formaldehyde and an enzymatic antigen retrieval step was performed. Samples were then incubated with ab14184 at a 1/100 dilution for 1 hour. The secondary used was an undiluted HRP conjugated goat anti-mouse polyclonal.

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Substance P staining at 1:1000 of the substancia gelatinosa (dorsal horn) of the guinea pig spinal cord using an HRP-conjugated secondary antibody and DAB.