how does it differ from conjugated antibodies? I used PE or FITC conjugated antibodies before but in this time, there is no available conjugated ab for this specific protein I want to measure. I tried to ask the net but I got confused instead. Thank you!
Hello Katharine,
If you have the antibody targeting your protein of interest, you can always get it conjugated with a reporter dye such as PE or FITC. Alternatively, you can use PE/FITC -conjugated secondary antibodies. Many companies sell such flow cytometry -validated secondary antibodies.
You can perform the staining using purified antibody. This will be a two-step procedure where you will 1st stain the cells with the primary antibody specific for the protein then use the secondary antibody conjugated with the fluorochrome. If your primary antibody is raised in mouse your secondary will be anti-mouse conjugated with PE/FITC. Also if your protein is intracellular you might have to perform the fixation and permeabilization step as well
I'd take Syed's advice and buy a fluorescent labelling kit. They are pretty much fool proof.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins