To visualize the subcellular localization of WT α/β and the various mutants, the different constructs were transfected into GNPTAB−/− HeLa cells using Lipofectamine 3000 (Life Technologies) according to the manufacturer’s protocol. 24 h post-transfection, the cells were fixed and the α/β subunits were detected with mouse anti-V5 monoclonal antibody (Life Technologies). The Golgi marker, GOLPH4, was detected with rabbit anti-GOLPH4 polyclonal antibody (Abcam, Cambridge, MA). The processed cells were mounted in ProLong® Gold antifade mounting medium (Life Technologies), and the images were acquired with an LSM880 confocal microscope (Carl Zeiss Inc., Peabody, MA). Images were analyzed by Image J software (Fiji).
Rabbit anti golph4 polyclonal antibody
Manufactured by Abcam
Rabbit anti-GOLPH4 polyclonal antibody is a reagent used in research applications. It is a purified antibody raised in rabbits against the GOLPH4 protein. GOLPH4 is a member of the golgin family of proteins and is involved in the structure and function of the Golgi apparatus.
Automatically generated - may contain errors
Lab products found in correlation
Prolong gold antifade mounting medium, by Thermo Fisher Scientific (2 mentions)
Lsm 880 confocal microscope, by Zeiss (2 mentions)
Mouse monoclonal anti v5 antibody, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
2 protocols using rabbit anti golph4 polyclonal antibody
1
Subcellular Localization of α/β Subunits
2
Subcellular Localization of α/β Subunits
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To visualize the subcellular localization of WT α/β and the S1-S3 mutant, the constructs were transfected into GNPTAB−/− HeLa cells using Lipofectamine 3000 according to the manufacturer’s protocol. Twenty-four hours post-transfection, the cells were fixed with 4% formaldehyde (Sigma-Aldrich) and the α/β subunits were detected with mouse anti-V5 monoclonal antibody (Life Technologies). The Golgi marker GOLPH4 was detected with rabbit anti-GOLPH4 polyclonal antibody (Abcam). The processed cells were mounted in ProLong Gold antifade mounting medium (Life Technologies), and the images were acquired with an LSM880 confocal microscope (Carl Zeiss). Images were analyzed using ImageJ software (Fiji).
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