1 × 106 treated Ishikawa cells were in a flat-bottom 24-well plate with cover glasses at the bottom. Cells were fixed with 4% v/v paraformaldehyde, permeabilized with 0.1% v/v Triton X-100 (Sigma, NJ, USA) and blocked them with 10% BSA in PBS. The cells were incubated with mouse anti-human GLS antibody (5 μg/ml, Abcam, MA, USA) at 4 °C overnight. Then the cells were incubated again with Alexa Fluor 488-conjugated anti-mouse secondary antibodies (1:1000; Cell Signaling Technology). The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI; Beyotime, Shanghai, China). The images were captured by using a confocal microscope (Leica, Munich, Germany).
Alexa fluor 488 conjugated anti mouse secondary antibodies
Manufactured by Cell Signaling Technology
Alexa Fluor 488-conjugated anti-mouse secondary antibodies are fluorescently labeled antibodies that recognize and bind to mouse primary antibodies. These secondary antibodies can be used in various immunodetection techniques, such as immunofluorescence and Western blotting, to visualize and detect target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Merck Group (1 mentions)
Confocal microscope, by Leica (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
γh2ax, by Merck Group (1 mentions)
α tubulin, by Abcam (1 mentions)
β actin, by Merck Group (1 mentions)
Total chk1, by Santa Cruz Biotechnology (1 mentions)
2 protocols using alexa fluor 488 conjugated anti mouse secondary antibodies
1
Immunofluorescence Staining of Glutaminase in Ishikawa Cells
2
Synthesis and Characterization of Ru1
Check if the same lab product or an alternative is used in the 5 most similar protocols
Ru1 was prepared using methods described by Bolger, et al.61 All NMR, mass spectroscopy and elemental analysis were in agreement with published data. Unless stated otherwise, all other chemicals were obtained from Sigma. Antibodies: p-Chk1 (Ser345) and p-Chk2 (Thr68) (Cell Signaling), γH2AX (Millipore), total Chk1 (Santa Cruz), total Chk2, α-tubulin (both Abcam) and β-actin (Sigma). AlexaFluor488-conjugated anti-mouse secondary antibodies were from Cell Signaling. DTPA-hEGF was prepared as reported previously.62 (link)
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