Human and mouse samples were fixed in formalin, dehydrated, embedded in paraffin, and sectioned. Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA) was used according to manufacturer’s instruction. Regarding human studies in Fig. 4 , skin samples from 5 healthy individuals and 5 patients with each following disease or disease subtype were used: dcSSc with an anti-topoisomerase I antibody, limited cutaneous SSc with an anti-centromere antibody, dcSSc with an anti-RNA polymerase III antibody, morphea, and eosinophilic fasciitis. As for mouse studies, the back skin and the right lower lobe of the lung were harvested. Skin thickness and cell counts were scored by S.N. and Y.A. in at least randomly chosen 5 high power fields of samples and the average number for each sample was used. Each section was examined independently in a blinded fashion. FITC-conjugated anti-rabbit (sc-2090, Santa Cruz) and Alexa Fluor 555-conjugated anti-goat (A21432; Life technologies, Carlsbad, CA, USA) were used for immunofluorescence studies as secondary antibodies at a 1/200 dilution.
Alexa fluor 555 conjugated anti goat a21432
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 555-conjugated anti-goat (A21432) is a fluorescently labeled secondary antibody. The Alexa Fluor 555 dye is conjugated to a goat anti-mouse IgG antibody, providing a tool for the detection and visualization of target proteins in various experimental applications.
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2 protocols using alexa fluor 555 conjugated anti goat a21432
1
Immunohistochemical Analysis of Skin and Lung
2
Immunohistochemical Analysis of Skin and Lung
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Human and mouse samples were fixed in formalin, dehydrated, embedded in paraffin, and sectioned. Vectastain ABC kit (Vector Laboratories, Burlingame, CA, USA) was used according to manufacturer’s instruction. Regarding human studies in Fig. 4 , skin samples from 5 healthy individuals and 5 patients with each following disease or disease subtype were used: dcSSc with an anti-topoisomerase I antibody, limited cutaneous SSc with an anti-centromere antibody, dcSSc with an anti-RNA polymerase III antibody, morphea, and eosinophilic fasciitis. As for mouse studies, the back skin and the right lower lobe of the lung were harvested. Skin thickness and cell counts were scored by S.N. and Y.A. in at least randomly chosen 5 high power fields of samples and the average number for each sample was used. Each section was examined independently in a blinded fashion. FITC-conjugated anti-rabbit (sc-2090, Santa Cruz) and Alexa Fluor 555-conjugated anti-goat (A21432; Life technologies, Carlsbad, CA, USA) were used for immunofluorescence studies as secondary antibodies at a 1/200 dilution.
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