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Cy3 conjugated goat anti mouse or anti rabbit

Manufactured by Jackson ImmunoResearch

Cy3-conjugated goat anti-mouse or anti-rabbit is a secondary antibody preparation that binds to mouse or rabbit primary antibodies. The Cy3 fluorescent label allows for the detection and visualization of target proteins in various immunoassay applications.

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2 protocols using cy3 conjugated goat anti mouse or anti rabbit

1

Histological and Immunostaining Analysis of Mouse Brains

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Mouse brains were fixed in 4% paraformaldehyde in PBS at 4 °C for overnight, embedded in paraffin and sectioned at 7 μm in the coronal or sagittal planes. For histological analyses, sections were rehydrated and stained with hematoxylin and eosin. For immunostaining, after rehydration, sections were incubated in boiling citrate buffer (10 mM, pH6.0) for 30 min for antigen retrieval. After three washes with PBS, primary antibodies (Supplementary Methods) were incubated with 5% normal goat or donkey serum in PBS at 4 °C for overnight. Sections were then washed in PBS and incubated with the secondary antibodies. Primary antibodies information can be found in Supplementary Information. For secondary antibodies, Alexa Fluor 488-conjugated goat anti-mouse, anti-rabbit or anti-chicken, 647 conjugated goat anti-mouse (Invitrogen), Cy3-conjugated goat anti-mouse or anti-rabbit (Jackson ImmunoResearch), and biotin-conjugated goat anti-rabbit (Vector Labs) were used. Images were acquired using Axioplan 2 (Carl Zeiss) and confocal microscopes (TCS SP5 and SP8, Leica) and analysed with LAS AF (Leica) and Photoshop (Adobe).
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2

Immunofluorescence Analysis of Cytoskeletal Proteins

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After 4 hours of culture, the culture medium was removed and unattached cells were washed away from the surface with PBS. Cells were fixed with 3 % paraformaldehyde in PBS for 30 min at 4 ⁰C, and washed three times with PBS. After permeabilizing with 0.1% Triton-X in PBS for 5 min at RT and blocking with DPBS/BSA 1 % at RT for 30 min, samples were incubated with a primary antibody for 1 hour at RT; the antibody used to analyze vinculin expression was antivinculin (mouse) (hVIN-1, Sigma-Aldrich) 1:400 in DPBS/BSA 1% and to analyze fibronectin expression was anti-fibronectin (rabbit) (polyclonal, Sigma-Aldrich) 1:400 in DPBS/BSA 1%.
Samples were washed twice with DPBS/Tween 20. A combination of secondary antibody (Cy3conjugated goat anti-mouse or antirabbit, respectively, Jackson ImmunoResearch) and phalloidin
(1:100) (BODIPY FL, Life Technology) was added and incubated by 1 hour at RT. Finally, after washing the samples with DPBS/Tween 20 three times, a mounting with Vectashield solution containing DAPI (Vector Laboratories) was performed.
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