AbC™ Total Antibody Compensation Bead Kit, 25 tests - FAQs

这是可能的。但是如果您的值超过100，可能需要查看你们用来采集数据的电压。

通过运行单色对照，有可能除去溢出到另一个荧光采集通道的荧光信号。

进行补偿时，你需要为自己使用的每种颜色参数准备单一染色样品（或补偿微球）。此外，我们建议您使用FMO（荧光扣除）对照。您可以使用减去一种颜色的其他所有颜色标记您的细胞或者微球来作为对照。为各种颜色准备FMO对照。这些对照对正确设置数据门限很有帮助。

理想情况下，每一个单独荧光团的荧光发射光谱都非常集中，窄峰，发射峰之间分隔的较开。现实中，有机染料和荧光蛋白有很宽的发射峰，必须考虑补偿（数据采集中或后）来矫正任何一个荧光团荧光信号的分配。补偿从重叠光谱中移除了荧光信号，使您知道所看见的信号就是目标荧光团发出的信号，所以非常重要。

补偿对照仅需未染色的细胞样品（或阴性对照）和单一颜色标记的样品——每管一个。如果您希望单色染色对照如您期望的那么亮,，那么可以使用我们的AbC总抗体补偿微珠试剂盒（货号A10513和A10497），这试剂盒是将抗体结合到微珠而非细胞上。

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.

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.

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.

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.

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.

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.

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.

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.