生物学医学各种试剂配方---很有用.doc

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1、Lysisbufferforwesternblot10*PBS20mlNP-40(nonidetp-40)1mlDOC*(sodiumdeoxycholate)0.5gSDS100mg100mMDTT1mlcactail5mlDW72.5mlTotal100ml1ml-2mlbuffer,whenhomogenizeHomogenizewithbigone*13000RPM,15minProteinassay4%PFA配方（1）60-70℃PBS（—）100ml（2）PFA4g（3）2NNaOHsoln100μL（如果是10NNaOH，就要用20μL[soln中H+的

2、量相同]）先把PBS放入60-70℃中，然后放入PFA和NaOH，直到PFA溶解后。配制好后放入4℃保存10XMopsbuffer0.4MMops0.1MNaacetate10mMEDTA.Na2PBS（—）[不含Ca+，Mg2+]Nacl8.0gKcl0.2gNa2HPO412H2O2.9gKH2PO40.2gmiliQUpto1L作用：相当于saline的作用CBB(考马斯亮蓝)染色液和脱色液配制ReagentmethanolAceticacidBrilliantblue1.Fixativebuffer(notusuallyusedingelinCBBstaini

3、ng)reagent1Lmethanol50mlAceticacid10mlMilliQ50ml2.CBBstainingbuffer(CBB染色液)(1L)reagentconcentration1Lmethanol40%(v/v)400mlAceticacid7%(v/v)70mlBrilliantblue0.25%(w/v)2.5gMilliQ530mlMethanol,aceticacidandmiliQweremixedfirstly,andbrilliantblue,then,wassolvedbymixedbuffer.Finally,CBBbuffer

4、couldstoreinR.T.3.Decolorizingbuffer(脱色液)(notusuallyusedinmembraneinCBBstaining)reagent1Lmethanol50mlAceticacid70mlMilliQ880ml常用缓冲溶液的配制方法1．甘氨酸–盐酸缓冲液（0.05mol/L）X毫升0.2mol/L甘氨酸+Y毫升0.2mol/LHCI，再加水稀释至200毫升pHXYpHXY2.02.42.62.85050505044.032.424.216.83.03.23.43.65050505011.48.26.45.0甘氨酸分子量=75.

5、07，0.2mol/L甘氨酸溶液含15.01克/升。 2．邻苯二甲酸–盐酸缓冲液（0.05mol/L）X毫升0.2mol/L邻苯二甲酸氢钾+0.2mol/LHCl，再加水稀释到20毫升pH(20℃)XYpH(20℃)XY2.254.0703.251.4702.42.62.83.055553.9603.2952.6422.0223.43.63.85550.9900.5970.263邻苯二甲酸氢钾分子量=204.23，0.2mol/L邻苯二甲酸氢溶液含40.85克/升 3．磷酸氢二钠–柠檬酸缓冲液pH0.2mol/LNa2HPO4（毫升）0.1mol/L柠檬酸（毫升）pH

6、0.2mol/LNa2HPO4（毫升）0.1mol/L柠檬酸（毫升）2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3010.6018.7617.8216.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.

7、2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量=14.98，0.2mol/L溶液为28.40克/升。Na2HPO4·2H2O分子量=178.05，0.2mol/L溶液含35.01克/升。C4H2O7·H2O分子量=210.14，0.1mol/L溶液为21.01克/升。 4．柠檬酸–氢氧化钠-盐酸缓冲液pH钠离子浓度（mol/L）柠檬酸（克）C6H8O7·H2O氢氧化