Immunofluorescence was performed as described [28 (link)]. The primary antibody was anti-α-Tubulin (Cell Signaling; mouse monoclonal; 1:2000) and the secondary antibody was Alexa488-conjugated anti-mouse IgG (Molecular Probes; 1:500). The slides were mounted in VectaShield fluorescence mounting medium containing 4, 6-diamidino-2-phenylindole (Vector Laboratories). Images were analyzed using a Zeiss confocal laser scanning microscope. Alternatively, cells were stained with α-tubulin antibody (1:50, mouse monoclonal; Abcam) followed by a CY5-conjugated secondary anti-mouse antibody (1:200; Abcam). The cover slips were mounted with Invitrogen Anti Fade Mounting Medium containing DAPI and imaged using a Zeiss AxioImager.Z1 microscope at 63x magnification.
Anti fade mounting medium
Manufactured by Zeiss
Anti Fade Mounting Medium is a lab equipment product designed to preserve and protect fluorescently labeled samples. It helps prevent the fading or quenching of fluorescent signals during microscopy and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa 488 conjugated anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Vectashield fluorescence mounting medium, by Vector Laboratories (1 mentions)
α tubulin antibody, by Abcam (1 mentions)
Anti α tubulin, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
2 protocols using anti fade mounting medium
1
Immunofluorescence Staining of α-Tubulin
2
Immunofluorescence Analysis of γ-H2AX
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were seeded in a cell slide, then washed with Phosphate Buffered Saline (PBST) after 24 h and fixed with 4% paraformaldehyde at 4°C for 30 min followed by 0.5% Triton X-100 20 min, and then incubated with blocking buffer (PBST containing 1% BSA) for 2 h. Next, cells were incubated with primary antibodies targeting γ-H2AX (phospho S139) (1:250 dilution) at 4°C overnight. Washed with PBST, cells were incubated with secondary antibodies (anti-rabbit Alexa 448-conjugated antibodies; 1:200 dilution; Beyotime Biotechnology) for 2 h at room temperature. Nuclei were stained with Hoechst (1:1000 dilution) for 2 min at room temperature. Cells were analyzed immediately with a confocal microscopy after adding antifade mounting medium (Zeiss).
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