Immunofluorescence analysis of CALD1 was performed using 70% confluent log phase MDA-MB-231 and MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Caldesmon Polyclonal Antibody (Product # PA5-17250) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image of MDA-MB-231 showing plasma membrane localization. Panel e represents merged image of MCF7 showing less expression of CALD1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Immunofluorescence analysis of CALD1 was performed using 70% confluent log phase MDA-MB-231 and MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Caldesmon Polyclonal Antibody (Product # PA5-17250) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image of MDA-MB-231 showing plasma membrane localization. Panel e represents merged image of MCF7 showing less expression of CALD1. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supplier Page from Thermo Fisher Scientific for Caldesmon Antibody