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After fixation in 0.3-percent glutaraldehyde and permeabilization with Triton X-100, an adherent culture of APM cells was treated with mouse anti-alpha-tubulin monoclonal primary antibodies followed by goat anti-mouse secondary antibodies (IgG) conjugated to Oregon Green 488. Mixed together with the secondary antibody was a phalloidin conjugate of Alexa Fluor 594, targeting the filamentous actin network. Nuclei were visualized by staining with Hoechst 33258 in Hanks' buffer for 30 minutes.
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