I have one experiment need flow cytometry, but we only have 1 non-conjugated antibody. Apart from buying a new antibody for flow cytometry. Could I use secondary antibodies in flow? How can I do during the staining?
Hello Shuo Wang
No, you cannot use non-conjugated secondary antibody in flow. Flow cytometry can be performed in two ways,
1) directly, using conjugated primary antibody or
2) indirectly, using a conjugated secondary antibody to bind an unconjugated primary antibody.
The indirect method would be more appropriate as indirect flow cytometry allows the choice of a wide range of probe molecules, enabling the user to match the desired probe with any primary antibody. Secondary antibody conjugates can improve a flow cytometry experiment by preserving the active site of the primary antibody, and by signal amplification.
In flow cytometry, a laser light is involved. Therefore, fluorescent detection with a secondary antibody conjugated to a fluorochrome is the requirement.
Best.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Methods
- Cloning