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1。我必须运行所有的标准和样品的重复？

是的，重复的运行，以监测测定的精度和提高检测结果的信心。

2。我必须在一次运行所有的样品吗？

不，每个试剂盒采用stripwell微孔板。这允许用户在不同的时间来分析不同数量的样品。

3。什么类型的可重复性的结果，得到与分析？

每一个工具都有一个包含一个典型数据的图表的手册。在运营商的任何变化，移液和洗涤技术，培养时间、温度、试剂盒的年龄可以引起变化的结果。每个用户都应该得到自己的标准曲线。

4。是否有可能储存的试剂比表示？

在不推荐使用条件下的部件组件的存储，以保证测试的正确性能。

5。我应该如何储存我的样品？

样品应保存在-20°C或更低的温度。长期储存，建议冻结他们在- 70余。

6。我可以修改协议吗？

BG的ELISA试剂盒进行了优化，提供*佳的结果。修改的格式或协议可能会给出错误的结果。

7。我可以使用一个不建议在试剂盒插入的样本类型吗？

该试剂盒已被验证在试剂盒插入的样品类型。未经检验的样品类型。为进一步信息的联系技术服务。

8。我的样本产生的值，在动态范围内的测定。我可以使用这些值吗？

建议只有在标准曲线的范围内下降的样本值。标准曲线的范围内的值一般是非线性的，这可能导致不正确的外推值。产生值高于标准的样品应为（进一步）稀释和重复。如果样品低于该测定的范围内，该样品被认为是不可检测的。

9。我每次都要运行一个空白或零标准吗？

是的，这些都是必需的计算，并反映任何微妙但显着的性能变化，从一天到一天，检测到检测。当排除特定的检测问题的来源时，它们也非常有帮助。

10。我能改变我在检测中使用的样本量吗？

不建议你改变卷由于BG工具被设计为*佳的性能在给定的体积

11。可以使用不同的组件组件吗？

每一个组件包含有特定批号的组件，以确保所有的组件都进行优化，以及与组件中的所有其他组件。对这些特定的地段进行质量检验。这是从来没有建议使用你自己的组件或组件从其他包或供应商。

12。我的标准曲线看起来很好，但我没有得到一个信号，我的样本，当我预期，为什么？

样品不包含被分析物。一个矩阵的效果可能是掩盖检测。确保推荐的稀释液在试剂盒插入后按规定进行。如果建议稀释，请检查是否正确进行稀释。过稀释会导致样品在标准曲线的范围内下降。

13。你怎么推荐我洗盘子？

如果您使用的是一个自动平板洗衣机，我们建议定期检查校准，并将该系统在洗涤前冲洗缓冲液冲洗。手动洗衣机也是一样的。一个中继器或洗瓶也可以使用。用户应小心，以确保所有的内容都是送气和板拍干的无尘纸。【大鼠水通道蛋白5(AQP-5)ELISA试剂盒】

14。我需要用一个盘子吗？

可靠的结果可以得到无板振动，但外径的一般会低于那些使用平板振动器。

15。为什么我要用波长校正450-570nm之间？

对于酶联免疫吸附试验，在双波长读的是做正确的光密度贡献的塑料以及，灯和光学波动。

16。如果我提我的样本，我还需要遵循推荐的稀释液试剂盒中插入了吗？

提取过程中所需的样本稀释量将受到影响的纯化和浓度的影响的协议使用。稀释或浓缩的金额将由*终用户决定。

17。什么是预期的分析物浓度，我应该找到？

一个给定的分析物的量可能会有所不同，不仅从物种的物种，但也组织和细胞来源。这个信息的*佳来源是目前的文献，很容易通过互联网在多个科学数据库访问。

18。我的光密度有点高（或较低），比那些在手册中，与我的试剂盒来了。为什么？

光学密度受影响

1.    Do I have to run all of my standards ａnd samples in duplicate?

Yes, the duplicates are run in order to monitor assay precision ａnd increase confidence in the assay results obtained.

2.    Do I have to run all of my samples at one time?

No, each kit uses stripwell microplate. This allows the user to analyse different numbers of samples at different times.

3.    What types of reproducible results are obtained with the assays?

Each kit comes with a manual containing a graph of typical data obtained. Any variation in operator, pipetting ａnd washing technique, incubation time or temperature, ａnd kit age can cause variation in result. Each user should obtain their own standard curve.

4.    Is it possible to store the reagents other than indicated?

Storage of the kit components under conditions other than indicated is not recommended in order to assure proper performance of the test.

5.    How should I store my samples?

Samples should be stored at -20oC or lower temperature. For long-term storage, it is recommended to freeze them at -70oC -80oC.

6.    Can I modify the protocol?

BG ELISA kits have been optimized to provide the best possible results. Modifying the format or protocol may give inaccurate ａnd wrong results.

7.    Can I use a sample type that is not recommended in the kit insert?

The kit has been validated for the sample types listed in the kit insert. Sample types other than those validated have not been tested. Contact Technical Service for further information.

8.    My samples generated values that were outside the dynamic range of the assay. Can I use these values?

It is recommended that only sample values that fall within the range of the standard curve be used. Values outside the range of the standard curve are generally non-linear, which can lead to incorrectly extrapolated values. Samples that generate values higher than the highest standard should be (further) diluted ａnd the assay repeated. If samples fall below the range of the assay, the sample is considered to be non-detectable. 【大鼠水通道蛋白5(AQP-5)ELISA试剂盒】

9.    Do I have to run a Blank or Zero Standards every time?

Yes, these are required for the calculations, ａnd reflect any subtle but significant performance changes from day to day ａnd assay to assay. They are also extremely helpful when troubleshooting the source of a particular assay problem.

10.  Can I alter the volume of sample I use in the assay?

It is not recommended that you alter the volumes since all BG kits are designed for optimal performance at the given volumes

11.  Can components from different kits be used?

Each kit contains components which have specific lot numbers to ensure that all of the components are performing optimally alone, as well as with all of the other components in the kit. QC testing is performed on these specific lots. It is never recommended to use your own components or components from other kits or vendors.

12.  My standard curve looked fine, but I didn’t get a signal in my sample when I expected to, why?

The sample may not contain the analyte. A matrix effect may be masking the detection. Ensure that the recommended dilution was followed as stated in the kit insert. If dilution was recommended, check to be sure that the dilution was performed properly. Over-dilution may cause the sample to fall below the range of the standard curve.

13.  How do you recommend I wash my plate?

If you are using an automated plate washer we recommend that the calibration be checked on a regular basis, ａnd that the system is flushed with the Plate Washing Buffer prior to washing. The same is true for a manual washer. A repeater or a wash bottle can also be used. The user should be careful to ensure that all of the contents are aspirated ａnd the plate tapped dry on lint-free paper.

14.  Do I need to use a plate shaker?

Reliable results can be obtained without a plate shaker, but the O.D.＇s will generally be lower than those obtained using a plate shaker.

15.  Why do I have to use wavelength correction between 450-570nm?

For the ELISA assay, reading at dual wavelengths is done to correct for the optical density contributed by the plastic well, the lamp ａnd optical fluctuations. 【大鼠水通道蛋白5(AQP-5)ELISA试剂盒】

16.  If I extract my sample, do I still need to follow the recommended dilutions given in the kit insert?

The amount of sample dilution needed after an extraction procedure will be affected by the effects of purification ａnd concentration in the protocol used. The amount of dilution or concentration will have to be determined by the end-user.

17.  What is the expected concentration of analyte that I should expect to find?

The amount of a given analyte may vary not only from species-to-species, but also between tissue ａnd cellular sources. The best source of this information is the current literature that is easily accessed through the Internet at multiple scientific databases.

18.  My optical densities were a little higher (or lower) than those in the manual that came with my kit. Why?

The optical density is affected by a number of physical conditions such as time ａnd temperature. We suggest that you shorten or lengthen the final incubation with substrate solution to compensate.

19.  What are the reasons for High Background?

1)     Improper Washing: Check volume of washing buffer reservoir ａnd make sure all recommended washing steps are performed.

2)     Contaminated Substrate: Make sure there is no contamination of the substrate with metal ions or oxidizing reagents, before use. Keep the extra substrate solution separately during the ELISA substrate development time. 【大鼠水通道蛋白5(AQP-5)ELISA试剂盒】

3)     Substrate exposed to light: Exposure to light may result in a blue color of the substrate. Keep solutions in the dark (vial) until ready to dispense into the plate.

4)     Wrong Incubation Times/Temperatures: Generally follow the test protocol regarding incubation times ａnd temperatures. However, if all wells are intensely ａnd equally colored with no intensity gradient observed in the standard dilution series, then it may be necessary to observe the substrate reaction as the color is developing, in order to stop the reaction sooner.

http://www.invitrogen.com/site/us/en/home.html

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