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AbC™ Anti-Rat/Hamster Bead Kit - FAQs

AbC™ Anti-Rat/Hamster Bead Kit - FAQs

View additional product information for AbC™ Anti-Rat/Hamster Bead Kit - FAQs (A10389)

6 product FAQs found

Does the AbC Anti-Rat/Hamster Bead Kit bind IgG2a kappa or IgG1 lambda rat with the same affinity?

Sorry, we have not analyzed this.

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.

I’ve been told that I need to compensate my data for flow cytometry analysis. What does that mean?

By running single-color controls, it is possible to remove signal of one fluorophore that spills over into the collection channel for another fluorophore.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

What kind of controls do I need for flow cytometry?

For compensation, you need to prepare a singly stained sample (or compensation beads) for each color parameter that you are using. In addition, we recommend that you use FMO (flow minus one) controls. These are controls in which you label cells or beads with every color in your panel, omitting one. Make one FMO control for each color. These controls are important for helping you properly set gates on your data.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Why should I worry about compensation in flow cytometry analysis?

In a perfect world, the fluorescence emission profile for each individual fluorophore would be a very intense, narrow peak, well separated from all other emission peaks. In reality, organic dyes and fluorescent proteins have broad emission peaks, and compensation must be employed (during or after data acquisition) to correctly assign fluorescence signal to each fluorophore. Compensation is important because it removes fluorescent signal from overlapping spectra so you know that the signal you see is only the signal from the fluorophore of interest.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

How do I make compensation controls for antibodies?

All that you need for compensation controls is an unstained sample of your cells (or negative control beads) and samples with each of your fluorophores - one per tube. You want the single-stained controls to be as bright as you expect your brightest sample to be. Antibodies can be bound to beads instead of cells using our AbC Total Antibody Compensation Bead Kit (Cat. Nos. A10513 and A10497).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.