OCT-embedded biopsy samples were cryosectioned and adhered to positively charged slides. Slides were fixed in 100% ethanol, blocked with 1% BSA and 5% milk, and stained with either mouse-anti IL-8 or rabbit-anti IL-6 primary antibodies (Abcam). Goat anti-mouse Cy3 or goat anti-rabbit Cy5 antibodies (Abcam) were used for secondary detection. Control sections were prepared by incubating tissue without primary antibody to confirm the absence of background staining. Images were captured with a fluorescent Zeiss Axio Imager microscope (Jena, Germany).
Goat anti mouse cy3
Manufactured by Abcam
Goat anti-mouse Cy3 is a secondary antibody conjugated with the fluorescent dye Cy3. It is designed to detect and visualize mouse primary antibodies in immunoassays and other applications.
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Lab products found in correlation
3 protocols using goat anti mouse cy3
1
Immunofluorescent Detection of Cytokines in OCT Biopsy
2
Quantifying Blood Vessel Density in Hypertrophic Scars
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Blood vessel density was assessed using co‐immunostaining for α‐smooth muscle actin (α‐SMA) and cluster of differentiation factor 31 (CD31) as previously described.23 Co‐staining was used due to the fact that immature blood vessels such as those in remodeling hypertrophic scars may not have full pericyte coverage that could be identified with α‐SMA alone.25 Biopsies taken at baseline representing normal skin (n = 4 Dc, n = 4 Yk) and at day 98 representing HTS (n = 4 Dc, n = 4 Yk) were sectioned and stained using primary antibodies for α‐SMA (Abcam, ab7817, 1:250, mouse) and CD31 (Biorbyt, orb10314, 1:500, rabbit) and secondary antibodies (goat anti‐mouse‐CY3 and goat anti‐rabbit‐CY5, Abcam). Two photos of the papillary dermis were taken at 10× magnification for each biopsy. Blood vessel density was calculated by counting the number of vessels and dividing by the area. Secondary antibody controls were stained in parallel and did not show nonspecific staining.
3
Immunohistochemical Localization of Podocalyxin
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Placental tissues were incubated with mouse podocalyxin (10 µg/mL) (R &D system, France) primary antibody. Incubation was performed overnight at 4 °C in PBS with 2% goat serum, 1% bovine serum albumin. Slides were then washed and incubated with a secondary antibody (Goat-antimouse Cy3, Abcam) for 1 hour at room temperature and counterstained with Hoechst 33342. Images were taken by Zeiss AxioVision microscope, and processed using AxioVision SE64 Rel. 4.9.1 software [28 (link)]. Negative controls were treated in an identical manner, except that podocalyxin antibody was replaced by PBS.
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