发布时间:2020-06-16 12:22 原文链接: ProcartaPlex免疫检测常见问题及解决方案

eBioscience公司Luminex液相蛋白芯片检测试剂——ProcartaPlex可以为您的生物学研究提供各种多因子检测方案,包括细胞因子、生长因子、趋化因子和其他蛋白。在选择试剂盒及操作时会遇到很多问题,现将常见问题及答复汇总如下,供您参考:

如果不小心将整个试剂盒保存在-20°C. 这个试剂盒还能再用吗?

I accidentally stored the whole kit at -20°C. Can I use it anyway?

A.    试剂盒中除微球外,其余都可以保存在-20oC。如果微球在0oC以下保存,会损伤微球使得微球不能再用了。

All  the components included in the ProcartaPlex kit are viable except the  bead mixture. Storage of the beads at temperatures below 0oC will damage  the beads and render them unusable.

如果我同时组合购买了多因子试剂盒和单因子试剂盒,还需要再买基础试剂盒吗?

Do I need to purchase a Basic Kit when combining Simplex Kits with a Multiplex Kit?

A.    不需要。

No,  when combining a Multiplex kit with Simplex Bead Sets, the Basic Kit is  not required. All the non target specific reagents which are included  in the Basic kit are components of our Multiplex kits. Only when you are  combining multiple Simplex kits (i.e. no Multiplex kit in the panel)  will you need to also purchase the Basic Kit.

我能否将小鼠基础试剂盒与人的单因子检测试剂盒搭配使用?

Can I use the ProcartaPlex Mouse Basic Kit with ProcartaPlex Human Simplex and vice versa?

A.    不能。基础试剂盒有种属特异性。不同种属的基础试剂盒之间不能交叉使用。

No,  the Basic Kits are specific to the species listed, i.e. the Mouse Basic  Kit can only be used with Mouse Simplex Bead Sets. The Mouse, Human,  Canine, Porcine, Rat and NHP ProcartaPlex Basic Kits are not  interchangeable.

多因子检测试剂盒不是同时购买的,可以搭配使用吗?

Can  I combine ProcartaPlex kits, which were not purchased on the same  order? For example: I purchased a kit 3 months ago and another kit  today. Can I combine them or do the kits have to be shipped on the same  order?

A.    可以的。只要确保同时检测的试剂盒的微球编号不重复,样本前期处理相同即可。

Yes,  you can combine ProcartaPlex kits from different orders to run a bigger  multiplex experiment. You will want to ensure that there is no overlap  of bead regions between the kits you are combining. Please refer to the  online ProcartaPlex Panel Configurator.

Luminex的微球能在普通流式细胞仪上检测吗?

Is it possible to measure Luminex beads on a flow cytometer?

A.    不行。检测ProcartaPlex (Luminex) 可以使用Luminex 100/200, MAGPIX, FLEXMAP 3D.

To  analyze ProcartaPlex (Luminex) plates, you will need access to a  Luminex machine such as a Luminex 100/200, MAGPIX, FLEXMAP 3D.

Luminex检测时,建议收集多少微球?

How many beads do you recommend we collect? And how do we setup the Luminex machine?

A.    推荐每种微球收集50-100个。

Please collect 50-100 beads per bead region as stated in the product manual.

ProcartaPlex酶标板在信号不损失的情况下,是否可以多次上机检测读板?

Can a finished ProcartaPlex plate be read multiple times on the Luminex instrument without a significant signal loss?

A.    可以的。

Yes,  it is possible to reread a finished ProcartaPlex plate without a loss  in the signal or the number of beads counted. Please note that the  Luminex instrument adds additional liquid to the wells with each  analysis. It is possible that the wells may become overfilled with fluid  after the third analysis. Hence, we do not recommend reading the  ProcartaPlex plates more than two times.

如果ProcartaPlex操作完成,不能及时上机检测。那我该如何保存?

I  have completed the ProcartaPlex assay but can’t run my samples  immediately. How long can I store the ProcartaPlex plate before analysis  on a Luminex instrument without a significant loss of signal?

A.    正常情况,我们建议立即上机检测;如果需要保存,建议在2-8oC保存。但是时间不能超过1天。

Ideally,  we recommend that you analyze the samples immediately. If you do need  to store your samples, overnight storage at 2-8°C shows no significant  signal loss. We do not recommend storing the ProcartaPlex plate longer  than 1 day.

洗涤步骤,我是否可以使用离心的方法操作?

Can I use a centrifuge for the washing steps in the assay?

A.    不可以。

No, we do not recommend centrifuging the magnetic beads for prolonged periods of time.

试剂盒中标准品溶解后,是否可以再保存使用?

Can I store reconstituted standard protein for later usage?

A.    不推荐。建议每次试验使用新的标准品。

The  recombinant standards included with the ProcartaPlex assay are one-time  use standards. Once reconstituted for use, we do not recommend storing  and reusing this recombinant standard. With each new experiment, we  recommend reconstituting a fresh unused vial of recombinant standard  protein.

标准品的初始溶解母液浓度从哪里得知?

Where can I find the stock concentration of the recombinant standards?

A.    请在随货收到的说明书中找到。

This information can be found on the Certificate of Analysis provided with the ProcartaPlex assay.

实验中,标准曲线和样本是否可以做单孔检测?

When preparing standards/samples, can I run them as single points?

A.    为了获得更好的结果准确性,我们建议样本和标准曲线孔至少做复孔。

For better accuracy, we recommend that you run samples/standards as duplicates at a minimum.

如果我不得不中止实验,我可以在哪步停止?

I  have to cut short the assay and can’t complete the assay. At which step  can we incubate the plates overnight with affecting the assay?

A.    按照说明书推荐,可以在加入标准品和样本后过夜反应。

As  stated in the product manual, after the addition of the standards and  samples, the plate can be stored overnight (Step 11.1.5 or 11.2.5).