Santa Cruz Biotechnology
cyclin A Antibody (H-432)
sc-751
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Browse Similar Products: cyclin A Immunofluorescence Antibodies
I used this cyclin A antibody to look at cell cycle using immunofluorescence. Previously I used the mouse monoclonal antibody from santa cruz, which also works well. However, this antibody is much easier to use in IF. Antibody has been used in western blots as well, very specific. Used at 1:2000 dilution for WB and 1:1000 dilution for IF. Can be used in either a 1 hour incubation, as the protocol below, or can be used at 4C overnight for IF. Both work very well. Protocol used: Cells were fixed in 3.5% paraformaldehyde for 15 minutes at room temperature.3 minutes in ice cold methanol. 5 minutes in PBS+0.5% Triton X5 minutes in PBS10 minutes in PBS+3%BSA. Antibody was incubated in PBS+1%BSA and coverslips incubated for 1 hour 2 - 5 minute washes in PBS Secondary antibodies from Jackson Laboratories (anti-rabbit) was incubated in PBS+1%BSA and coverslips 2 - 5 minute washes in PBSMounted coverslips using ProLong Gold
Immunofluorescence
RPE-1 cells
1:2000 in PBS+1% BSA 1 hour room temp
PBS+3%BSA
1:1000 in PBS+1%BSA for 1 hour
None
Fluorescence imaging was done on a 60X objective
Cells were stained against cyclin A (red) and costained with ACA (green) and DAPI (blue) and imaged using a 60X objective on a Nikon micoroscope. Asynchronous population of cells are seen in the image.
Works well
Nothing
Very good antibody against cyclin A