Mon Oct 20, 2014 6:05 pm by mleipold
HI Chad,
I have used BD's anti-mouse Ig K capture beads (cat #552843) pretty successfully for single-metal antibody bead controls.
This has worked with the vast majority of my MAXPAR-labeled antibodies. Out of 33 or so tested, I think I've only had 2 consistently give issues: very dim or no signal on the beads, while still staining cells properly. I haven't bothered to track down the issue (eg, no kappa chain).
When I do these experiments, I run one bead-antibody at a time. Therefore, just gating on a high-M-metal signal is usually sufficient. I do make sure to do a number of washes, though.
I should mention: even with this bead data, we haven't been able to make a "compensation" matrix in FlowJo. Whether this is a math problem because of all the zeroes in the negative data, we don't know. But FlowJo doesn't make much, if any adjustment, with compensation on or off. This includes cases where we *know* there's an isotopic spillover (eg, Dy163 impurity in both Dy162 and Dy164 stocks).
If anyone has suggestions or comments regarding that, please chime in.
Mike