5μm sections from FFPE tissue were deparaffinized using a xylene-ethanol gradient, rehydrated, and rinsed in phosphate-buffered saline (PBS). Antigen retrieval was performed by immersion in a modified pH 6.0 citrate buffer for 20 minutes in a steamer. Slides were cooled and rinsed in PBS. Tissue sections or transfected cells were fixed with 1:1 acetone:methanol for 10 minutes at −20°C. Blocking was performed with 10% donkey serum/1% BSA for 1 hour at room temperature. Slides were incubated 1 hour with primary antibody, washed 3 times with 1X PBS, incubated 1 hour with secondary antibody, and again washed 3 times with 1X PBS before mounting with Mowiol/1% n-propyl gallate (Polysciences, Warrington, PA). Primary antibodies: 1:100 rabbit anti-Cx43, 1:100 goat anti-HA, 1:70 mouse anti-GM130 (ab11370, ab9134, ab169276; Abcam, Cambridge, England). Secondary antibodies: 1:10000 Cy3 donkey anti-rabbit IgG, 1:10000 Cy3 donkey anti-goat IgG, 1:200 Cy2 donkey anti-mouse IgG (711-165-152, 711-165-003, 711-225-150; Jackson ImmunoResearch, West Grove, PA).
Cy2 donkey anti mouse igg
Manufactured by Jackson ImmunoResearch
Cy2 donkey anti-mouse IgG is a secondary antibody conjugated with the Cy2 fluorescent dye. It is designed to detect and visualize mouse immunoglobulin G (IgG) in various immunoassays and imaging applications.
Automatically generated - may contain errors
2 protocols using cy2 donkey anti mouse igg
1
Immunofluorescence Staining of FFPE Tissue
2
Immunofluorescence Staining of FFPE Tissue
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5μm sections from FFPE tissue were deparaffinized using a xylene-ethanol gradient, rehydrated, and rinsed in phosphate-buffered saline (PBS). Antigen retrieval was performed by immersion in a modified pH 6.0 citrate buffer for 20 minutes in a steamer. Slides were cooled and rinsed in PBS. Tissue sections or transfected cells were fixed with 1:1 acetone:methanol for 10 minutes at −20°C. Blocking was performed with 10% donkey serum/1% BSA for 1 hour at room temperature. Slides were incubated 1 hour with primary antibody, washed 3 times with 1X PBS, incubated 1 hour with secondary antibody, and again washed 3 times with 1X PBS before mounting with Mowiol/1% n-propyl gallate (Polysciences, Warrington, PA). Primary antibodies: 1:100 rabbit anti-Cx43, 1:100 goat anti-HA, 1:70 mouse anti-GM130 (ab11370, ab9134, ab169276; Abcam, Cambridge, England). Secondary antibodies: 1:10000 Cy3 donkey anti-rabbit IgG, 1:10000 Cy3 donkey anti-goat IgG, 1:200 Cy2 donkey anti-mouse IgG (711-165-152, 711-165-003, 711-225-150; Jackson ImmunoResearch, West Grove, PA).
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