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1。我必須運(yùn)行所有的標(biāo)準(zhǔn)和樣品的重復(fù)？

是的，重復(fù)的運(yùn)行，以監(jiān)測測定的精度和提高檢測結(jié)果的信心。

2。我必須在一次運(yùn)行所有的樣品嗎？

不，每個(gè)試劑盒采用stripwell微孔板。這允許用戶在不同的時(shí)間來分析不同數(shù)量的樣品。

3。什么類型的可重復(fù)性的結(jié)果，得到與分析？

每一個(gè)工具都有一個(gè)包含一個(gè)典型數(shù)據(jù)的圖表的手冊。在運(yùn)營商的任何變化，移液和洗滌技術(shù)，培養(yǎng)時(shí)間、溫度、試劑盒的年齡可以引起變化的結(jié)果。每個(gè)用戶都應(yīng)該得到自己的標(biāo)準(zhǔn)曲線。

4。是否有可能儲存的試劑比表示？

在不推薦使用條件下的部件組件的存儲，以保證測試的正確性能。

5。我應(yīng)該如何儲存我的樣品？

樣品應(yīng)保存在-20°C或更低的溫度。長期儲存，建議凍結(jié)他們在- 70余。

6。我可以修改協(xié)議嗎？

BG的ELISA試劑盒進(jìn)行了優(yōu)化，提供*佳的結(jié)果。修改的格式或協(xié)議可能會給出錯(cuò)誤的結(jié)果。

7。我可以使用一個(gè)不建議在試劑盒插入的樣本類型嗎？

該試劑盒已被驗(yàn)證在試劑盒插入的樣品類型。未經(jīng)檢驗(yàn)的樣品類型。為進(jìn)一步信息的聯(lián)系技術(shù)服務(wù)。

8。我的樣本產(chǎn)生的值，在動態(tài)范圍內(nèi)的測定。我可以使用這些值嗎？

建議只有在標(biāo)準(zhǔn)曲線的范圍內(nèi)下降的樣本值。標(biāo)準(zhǔn)曲線的范圍內(nèi)的值一般是非線性的，這可能導(dǎo)致不正確的外推值。產(chǎn)生值高于標(biāo)準(zhǔn)的樣品應(yīng)為（進(jìn)一步）稀釋和重復(fù)。如果樣品低于該測定的范圍內(nèi)，該樣品被認(rèn)為是不可檢測的。

9。我每次都要運(yùn)行一個(gè)空白或零標(biāo)準(zhǔn)嗎？

是的，這些都是必需的計(jì)算，并反映任何微妙但顯著的性能變化，從一天到一天，檢測到檢測。當(dāng)排除特定的檢測問題的來源時(shí)，它們也非常有幫助。

10。我能改變我在檢測中使用的樣本量嗎？

不建議你改變卷由于BG工具被設(shè)計(jì)為*佳的性能在給定的體積

11�？梢允褂貌煌慕M件組件嗎？

每一個(gè)組件包含有特定批號的組件，以確保所有的組件都進(jìn)行優(yōu)化，以及與組件中的所有其他組件。對這些特定的地段進(jìn)行質(zhì)量檢驗(yàn)。這是從來沒有建議使用你自己的組件或組件從其他包或供應(yīng)商。

12。我的標(biāo)準(zhǔn)曲線看起來很好，但我沒有得到一個(gè)信號，我的樣本，當(dāng)我預(yù)期，為什么？

樣品不包含被分析物。一個(gè)矩陣的效果可能是掩蓋檢測。確保推薦的稀釋液在試劑盒插入后按規(guī)定進(jìn)行。如果建議稀釋，請檢查是否正確進(jìn)行稀釋。過稀釋會導(dǎo)致樣品在標(biāo)準(zhǔn)曲線的范圍內(nèi)下降。

13。你怎么推薦我洗盤子？

如果您使用的是一個(gè)自動平板洗衣機(jī)，我們建議定期檢查校準(zhǔn)，并將該系統(tǒng)在洗滌前沖洗緩沖液沖洗。手動洗衣機(jī)也是一樣的。一個(gè)中繼器或洗瓶也可以使用。用戶應(yīng)小心，以確保所有的內(nèi)容都是送氣和板拍干的無塵紙。【大鼠1,4,5-三磷酸肌醇(IP3)ELISA試劑盒】

14。我需要用一個(gè)盤子嗎？

可靠的結(jié)果可以得到無板振動，但外徑的一般會低于那些使用平板振動器。

15。為什么我要用波長校正450-570nm之間？

對于酶聯(lián)免疫吸附試驗(yàn)，在雙波長讀的是做正確的光密度貢獻(xiàn)的塑料以及，燈和光學(xué)波動。

16。如果我提我的樣本，我還需要遵循推薦的稀釋液試劑盒中插入了嗎？

提取過程中所需的樣本稀釋量將受到影響的純化和濃度的影響的協(xié)議使用。稀釋或濃縮的金額將由*終用戶決定。

17。什么是預(yù)期的分析物濃度，我應(yīng)該找到？

一個(gè)給定的分析物的量可能會有所不同，不僅從物種的物種，但也組織和細(xì)胞來源。這個(gè)信息的*佳來源是目前的文獻(xiàn)，很容易通過互聯(lián)網(wǎng)在多個(gè)科學(xué)數(shù)據(jù)庫訪問。

18。我的光密度有點(diǎn)高（或較低），比那些在手冊中，與我的試劑盒來了。為什么？

光學(xué)密度受影響

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1,4,5-三磷酸肌醇(IP3)ELISA試劑盒】

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1,4,5-三磷酸肌醇(IP3)ELISA試劑盒】

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1,4,5-三磷酸肌醇(IP3)ELISA試劑盒】

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.

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