We randomly chose 50 tissue specimens for FGF
staining according to higher and lower than the estimated
relative quantification (RQ) median (12.39) for FGF10.
The first paraffin separation was done at 55°C, then, after
20 minutes, three washes with xylene. Next, the samples
were rehydrated in ethanol concentrations of 100%, 95%,
and 80%. Subsequently, 3% hydrogen peroxide was
applied to block any endogenous peroxidase activity.
The prepared slides were heated in citrate acidic buffer
(pH=6.0) in an oven for 10 minutes through antigen
retrieval. Primary antibody (1:100) incubation at 4°C
was done for two hours, then each slide was incubated
for 30 minutes in biotin labelled with secondary antibody
(Abcam, ab80064, Cambridge, UK) and tracked with
incubation in streptavidin-peroxidase (Dakocytomation,
Inc., CA, USA). The Diaminobenzidine substrate was
added to the slides, and they were visualized under a
microscope. Finally, the cutting segments were stained
with haematoxylin and dehydrated. For checking the
negative control, the intended segment was incubated
with phosphate-buffered saline (PBS) instead of primary
antibody, followed by the rest of the previously mentioned
procedures. The positive control was brain-skin tissue
segments, which had high FGF10 expression (1 (link)).
staining according to higher and lower than the estimated
relative quantification (RQ) median (12.39) for FGF10.
The first paraffin separation was done at 55°C, then, after
20 minutes, three washes with xylene. Next, the samples
were rehydrated in ethanol concentrations of 100%, 95%,
and 80%. Subsequently, 3% hydrogen peroxide was
applied to block any endogenous peroxidase activity.
The prepared slides were heated in citrate acidic buffer
(pH=6.0) in an oven for 10 minutes through antigen
retrieval. Primary antibody (1:100) incubation at 4°C
was done for two hours, then each slide was incubated
for 30 minutes in biotin labelled with secondary antibody
(Abcam, ab80064, Cambridge, UK) and tracked with
incubation in streptavidin-peroxidase (Dakocytomation,
Inc., CA, USA). The Diaminobenzidine substrate was
added to the slides, and they were visualized under a
microscope. Finally, the cutting segments were stained
with haematoxylin and dehydrated. For checking the
negative control, the intended segment was incubated
with phosphate-buffered saline (PBS) instead of primary
antibody, followed by the rest of the previously mentioned
procedures. The positive control was brain-skin tissue
segments, which had high FGF10 expression (1 (link)).
