elisa标准曲线制作方法

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1、ELISA标准曲线制作方法一般而言，我们拟合ELISA标准曲线选用比较经典的CurveExpert1.3或者CurveExpert1.4软件。现在我们以CurveExpert1.4为例，对ELISA标准曲线绘制方法进行详述。Generallyspeaking,thestandardfittingcurveofELISAisbasedontheclassicsoftwareofCurveExpert1.3orCurveExpert1.4.NowwewillusetheCurveExpert1.4toillustratehowtoconstructtheELISAstandard

2、curve.1.点击CurveExpert1.4，打开应用程序，截图界面如下：ClickCurveExpert1.4,andthenopentheapplicationprograms.Youwillseethefollowingscreenshot.2.在X轴输入标准品的OD值，Y轴输入相应的标准品的浓度，截图界面如下：InputthevalueofODontheX-axisagainsttheconcentrationofsamplesontheY-axis.Thefollowingscreenshotwillappearlikethis.3.单击上图界面中的图标，出现如

3、下界面Clicktheiconshowedontheabovescreenshotandyouwillseethefollowingpicture.4.单击上图界面中的ALLOFF按钮，出现如下界面Clickthe‘AllOff’button,youcanseethefollowingscreenshot.3.我们选择SigmoidalModels这种曲线拟合模型进行ELISA标准曲线的拟合，点击OK，出现下图界面：WeselecttheSigmoidalModelstofittheELISAstandardcurve.Click‘OK’,andyouwillseethefo

4、llowingscreenshot.注意：上图界面中的r值越高，即当r值越接近1时，所拟合的曲线就越好，所得到的数值可靠性也就也高。Pleasenotethatthehigherther-valueintheabovepicture(orthecloserther-valueisto1),themorereliablevaluesyouwillget.3.在上图空白界面，按[Ctrl]键+[L]键，出现如下界面PleasepresstheCtrlplusLintheblankpartoftheabovepicture.Theinterfacewillappearlikethi

5、s.在上图中输入相应的OD值（X值）后，点击Calculate即可计算出待测物的浓度（Y值），待测物稀释了多少倍就在所测值的基础上乘以稀释倍数即最终浓度。AfterinputtingthecorrespondingOD-value(x-value),clickthebutton“Calculate”.Theconcentration(Y-value)ofsamplescanbemeasured.Thefinalconcentrationcanbeobtainedbymultiplyingthethemeasuredfigurewiththetimesthesampleshav

6、ebeendiluted.3.点击所绘制的曲线图中左上方按钮，可以得到如下ELISA曲线拟合方程Clickthebuttononthetopleftcornerinthepicture.ThenyoucangettheELISAcurvefittingequationasfollows.3.点击上图中Copy即可得到上图界面中的曲线方程数据LogisticModel:y=a/(1+b*exp(-cx))CoefficientData:a=-1.19763443774E+004b=-8.45403498052E+001c=1.30213993104E+000Clickthebu

7、tton“copy”andyouwillgetthefiguresforcurveequationshowedintheabovescreenshot.LogisticModel:y=a/(1+b*exp(-cx))CoefficientData:a=-1.19763443774E+004b=-8.45403498052E+001c=1.30213993104E+000