CYTOSKELETON, INC
Rhodamine Phalloidin
PHDR1
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Our aim was to examine F-actin staining in cardiomyocytes. We transfected adult cardiomyocytes with the scramble and target specific siRNA. It was analyzed with F-actin specific antibody. Antibody is easy to optimize and binds specifically to F-actin and does therefore stain the actin stress fibers in the cell
Immunofluorescence
Rat cardiomyocyte
Rhodamine Phalloidin (1:200 dilution), Overnight at 4 degree
3% BSA for 30 mins
Actin-stain™ 555 cells were incubated with a TRITC-conjugated antibody (1:5000)
N/A
Confocal microscopy was performed with a Zeiss LSM 510 Pascal or Zeiss LSM710 confocal microscopes.
I proceed by fixing the cell (on glass coverslips) with 4%PFA (in 1x PBS) (20 min at RT) then permeabilized with 0.5% Trito-X100 (in 1X PBS) (at RT for 30 mins) followed by blocking with 3% BSA in 1x PBS . Incubate the cells with 1;200 phallodind actin made in 1XPBS for O/N at RT. Washed 3 times, then added TRITC-conjugated secondary antibody (1:5000). Washed 3 times with 1X PBS. Mounting on glass slide with antiquenching agent, seal with the nail paint.I observed a very good staining on actin fibers.
very good
None
This is one of best antibody for actin staining. I recommend strongly.