I need to label my tissue section with two mouse primary antibodies. Do I have to make direct conjugates of the antibodies to use them?
Check the isotype of the two antibodies to see if they are different. If so, we offer isotype-specific goat anti-mouse secondary antibodies that can be used together with minimal cross-reactivity. If the two mouse antibodies are of the same isotype, then you would need to make direct conjugates, such as with our antibody labeling kits or APEX, and SiteClick kits.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
After adding the antibody to the AbC Total Antibody Compensation beads, there does not appear to be any signal. What may have caused this?
Here are some tips to follow:
- Make sure that the beads were never frozen.
- Avoid mixing by vortexing too long (no longer than 10 seconds) or sonication as this may promote denaturation of the protein on the surface of the beads.
- Use the product within the warranty period (one year).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Can the AbC Total Antibody Compensation Bead Kit bind Fab dimers?
Yes, the AbC Total Antibody Compensation Bead Kit can bind Fab dimers as long as the Fab dimers are derived from either mouse, rat, hamster, or rabbit species.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The AbC Total Antibody Compensation Bead Kit accommodates only mouse, rat, hamster, and rabbit antibodies. What can I use if my antibody is not derived from any of these species?
As an alternative, you may covalently attach your antibody to colorless, 5 µm Aldehyde/Sulfate Latex Beads (Cat. No. A37306). The aldehyde moiety on the bead surface directly binds with primary amines on the surface of the antibodies in simple buffers such as PBS (pH 7.4). Make sure that the buffer does not contain any primary amines (i.e., do not use Tris or glycine buffers). To avoid making bead-antibody-bead-antibody multimers, add excess antibody relative to the beads.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
I am performing flow cytometry. My compensation matrix has values over 100. That's not possible is it?
It is possible. But if you have a lot of values over 100, you probably need to look at the voltages that you are using for your data collection.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.