Fusion灰度分析半定量WesternBlot条带步骤

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1、Fusion灰度分析半定量WesternBlot条带步骤1.打开图片分析图片处理分析FUSION分子呈◎llllllllllllllllllll33788>2•Q备注：此时可以调节图片灰度情况，但是对最终的结果影响不大2L分析嬪*岂片介理I■m!Siiiiiiiiiiiiiiiiiiii看到一下界面：2、圈出要分析的条带方法（1）整条一起圈出来，用于连续条带，条带位置差距不大，宽度差距不大的51回1(13團

2、ArviModule方法（2）每个条带分别圈出来■AnjModue.TempfileWindowdefwwtionSpotq

3、Copy•/——3、下一步An>Module-TempKle1WindowdrflmttonNewlaneoIIG9ifs4.去背景IoIfE)If£3IAn^Module.TempFfle]Auloff去背景后5、下一步出现一下界面：AruModuleTempRe"o'O「旨Spotseparation•LaneAIhresholdvalue:U31120-I»72NumberVol%VolumeHeightAreaNol121158.939.M724.9161No2102927.594.52451.U3NoJ12M99.28135226.9131No491926.7独

4、279M4MNo$167S0UJ59.WOX.9742NO*106937.190.71713.9442No770555.206.29617.9$6IMO86W110.M4No976375.635.3S810.2702No1076265,627.46812.497141m6、分隔条带：对软件自动识别的条带进行修正，删除不必要的条带(Deleteseparation)或增加新的条带(Addseparation)IIrabonf>apSig1耐Spotseparation•LaneAThresholdvalue:0NumberVol%VolumlNol1211S8.95

5、No2

6、102927.5SINoJ125899.2(1No491S26.7!

7、NoSM75012.3!

8、NO6106957.5S

9、No7705$S.2C

10、NoS*0914.4slNo974S75.6:1No10762656：］14»1No95321No11No216290No314.233No411860No57.573No87038No10NO12qModfythespotseparabonsUnenumber:AddseparatorDeteteseparabon7、指定作为100%参考量的参考条带8、输出结果■AmModule.TempFSeo卫宣9X结果分析相对灰度计算：目

11、的条带的Volume/内参的VolumeMIF.rggcondgBCSJumbVol箱VolumeHeightArMNO11059883,0111.0652.544No2120555435.0248.47Q2.703NO314676.613.05010.8242.809No407054375.3117.0722650No5IM807.1831.5032.438No6174S17.872.23811.4682.968No?150047.170.47010.88：2703NO8123085.549.2287.8272.862GAPDHNumbtfvol・ConN0111406C

12、uNO2115147EPANO310232ECSNo412068ConNO50020CuNO610857TEPANO76651ECSNO85392Volume5.5117264•smdo16353688C97545J5a2H^ghtArM10.68S2.77312.8622.38710.0522.44412.801255510.0322.45110.8522.4448.0202AU6.9132.5S5HIF-1O/GAPDH01602C589609S452107813482344100-5613*35302072535C215123350922487211042146622

13、73810.三次重复实验，归一HIF-la/GAPDK3组减丟Con0.16020500.2072541.615030660830000.2424310.313632.443943Cu0.08452111.5123352.883091.4808252.288544.3628320TEPA二134823442.2487211.887542.0402143.4028728563215BCS二0.75610742.1466231.9002811443153.248372.889218411.统计软件进行统计