Approximately thirty hours following transient transfected of either pCMV-Flag-HA-SLC25A51 or pCMV-Flag-HA-SLC25A52, HeLa cells seeded on coverslips were fixed using 4% paraformaldehyde (Electron Microscopy Sciences #15710)/PBS for 15 minutes at room temperature. Fixed cells were then washed in PBS, blocked, and permeabilized for 1 hour at room temperature in 5% normal goat serum/0.3% Triton X-100/PBS. Primary antibodies were diluted and incubated with cells overnight at 4ºC in 1% BSA/0.3% Triton X-100/PBS. Rabbit anti-Flag (Cell Signaling Technologies #14793, 1:500); mouse anti-MTC02 (Abcam ab79479, 1:40). After washing, secondary antibodies were similarly diluted and incubated with cells for 1 hour at room temperature. Goat anti-Mouse IgG - Alexa Fluor 488 (Invitrogen A-11001, 1:1000); Goat anti-Rabbit IgG - Alexa Fluor 568 (Invitrogen A-11036, 1:1000). Following 3X PBS washes, cells were mounted with Vectashield Hardset w/ DAPI (Vector Labs, H-100). 0.11 μm optical slices were imaged using a Yokogawa W2 spinning disk confocal setup that includes 100mW 488 nm and 565 nm lasers, a 100X Olympus objective, and a Photonics Prime 95B sCMOS camera.
Mouse anti mtc02
Manufactured by Abcam
Mouse anti-MTC02 is a monoclonal antibody that specifically recognizes the mitochondrial cytochrome c oxidase subunit II (MTC02) protein. This antibody can be used for the detection of MTC02 in various applications such as Western blotting, immunohistochemistry, and flow cytometry.
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2 protocols using mouse anti mtc02
1
Subcellular Localization of SLC25A51/52
2
Subcellular Localization of SLC25A51/52
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Approximately thirty hours following transient transfected of either pCMV-Flag-HA-SLC25A51 or pCMV-Flag-HA-SLC25A52, HeLa cells seeded on coverslips were fixed using 4% paraformaldehyde (Electron Microscopy Sciences #15710)/PBS for 15 minutes at room temperature. Fixed cells were then washed in PBS, blocked, and permeabilized for 1 hour at room temperature in 5% normal goat serum/0.3% Triton X-100/PBS. Primary antibodies were diluted and incubated with cells overnight at 4ºC in 1% BSA/0.3% Triton X-100/PBS. Rabbit anti-Flag (Cell Signaling Technologies #14793, 1:500); mouse anti-MTC02 (Abcam ab79479, 1:40). After washing, secondary antibodies were similarly diluted and incubated with cells for 1 hour at room temperature. Goat anti-Mouse IgG - Alexa Fluor 488 (Invitrogen A-11001, 1:1000); Goat anti-Rabbit IgG - Alexa Fluor 568 (Invitrogen A-11036, 1:1000). Following 3X PBS washes, cells were mounted with Vectashield Hardset w/ DAPI (Vector Labs, H-100). 0.11 μm optical slices were imaged using a Yokogawa W2 spinning disk confocal setup that includes 100mW 488 nm and 565 nm lasers, a 100X Olympus objective, and a Photonics Prime 95B sCMOS camera.
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