Tissue samples analyzed in this study were perfused with PBS and fixed with 10% Neutral Buffered Formalin (VWR 16004–128). Fixed tissues were paraffin embedded, sectioned and stained using standard histologic techniques. Immunofluorescence slides were deparaffinized, rehydrated and retrieval was performed in citrate buffer, pH 6.0 (Abcam AB93678) in a programmable decloaker (Biocare DC2012). Slides were permeabilized with PBS + 0.05% Tween-20 (Sigma P9416) and blocked with Sea Block (Thermo 37527). Primary antibodies used included mouse anti-CD31 (Abcam AB187377), rabbit anti-Collagen I (Abcam AB34710), rabbit anti-LYVE1 (Abcam AB33682), and mouse anti-C4D (Abcam AB90804). Secondary antibodies used were goat anti-rabbit Alexa Fluor 488 (Thermo A11078), goat anti-mouse Alexa Fluor 488 (Thermo A11029), and goat anti-rabbit Alexa Fluor 555 (Thermo A21429). Slides were stained with 4’,6-diamidino-2-phenylindole (Thermo D1306) diluted 1:200 in PBS and mounted using ProLong Antifade Mountant (Thermo P36961). Fluorescence slides were imaged on a Zeiss Axioskop 40 and H&E slides were imaged on an Accuscope 3012.
Dc2012
Manufactured by Biocare Medical
The DC2012 is a laboratory centrifuge designed for general-purpose applications. It features a fixed-angle rotor that can accommodate various sample containers. The centrifuge is capable of achieving a maximum speed of 12,000 rpm and a maximum relative centrifugal force (RCF) of 17,000 x g. The DC2012 is a reliable and versatile piece of equipment for separating and processing samples in a laboratory setting.
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2 protocols using dc2012
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Immunofluorescence Staining of Paraffin-Embedded Tissues
2
Immunofluorescence Staining of Paraffin-Embedded Tissues
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Tissue samples analyzed in this study were perfused with PBS and fixed with 10% Neutral Buffered Formalin (VWR 16004–128). Fixed tissues were paraffin embedded, sectioned and stained using standard histologic techniques. Immunofluorescence slides were deparaffinized, rehydrated and retrieval was performed in citrate buffer, pH 6.0 (Abcam AB93678) in a programmable decloaker (Biocare DC2012). Slides were permeabilized with PBS + 0.05% Tween-20 (Sigma P9416) and blocked with Sea Block (Thermo 37527). Primary antibodies used included mouse anti-CD31 (Abcam AB187377), rabbit anti-Collagen I (Abcam AB34710), rabbit anti-LYVE1 (Abcam AB33682), and mouse anti-C4D (Abcam AB90804). Secondary antibodies used were goat anti-rabbit Alexa Fluor 488 (Thermo A11078), goat anti-mouse Alexa Fluor 488 (Thermo A11029), and goat anti-rabbit Alexa Fluor 555 (Thermo A21429). Slides were stained with 4’,6-diamidino-2-phenylindole (Thermo D1306) diluted 1:200 in PBS and mounted using ProLong Antifade Mountant (Thermo P36961). Fluorescence slides were imaged on a Zeiss Axioskop 40 and H&E slides were imaged on an Accuscope 3012.
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