标准缓冲液的配制方法

1、标准缓冲液的配制及常用数据一、标准缓冲液pH值与温度对照表 温度四草酸氢钾0.05M邻苯二甲酸氢钾0.05M 混合磷酸盐0.025M硼砂0.01M51.674.006.959.39101.674.006.029.33151.674.006.909.28201.684.006.889.23251.684.006.869.18301.684.016.859.14351.694.026.849.11 二、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0.2 mol/L甘氨酸+Y毫升0.2 mol/L HCI，再加水稀释至200毫升pHXYpHXY

2、2.02.42.62.85050505044.032.424.216.83.03.23.43.65050505011.48.26.45.0甘氨酸分子量 = 75.07，0.2 mol/L甘氨酸溶液含15.01克/升。 2邻苯二甲酸盐酸缓冲液（0.05 mol/L）X毫升0.2 mol/L邻苯二甲酸氢钾 + 0.2 mol/L HCl，再加水稀释到20毫升pH(20XYpH(20XY2.22.42.62.83.0555554.0703.9603.2952.6422.0223.23.43.63.855551.4700.9900.5970.263邻苯二甲酸氢钾分子量 = 204.23，0.

3、2 mol/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/L Na2HPO4（毫升）0.1mol/L柠檬酸（毫升）pH0.2mol/L Na2HPO4（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

4、.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量 = 14.98，0.2 mol/L溶液为28.40克/升。Na2HPO4-2H2O分子量 = 178.05，0.2 mol/L溶液含35.01克/升。C4H2O7·H2O分子量 = 210.14，0.1 mol/L溶

5、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氢氧化钠(克NaOH 97%盐酸（毫升）HCl（浓）最终体积（升）2.23.13.34.35.35.86.50.200.200.200.200.350.450.3821021021021024528526684838383144186156160116106456810512610101010101010     使用时可以每升中加入1克克酚，若最后pH值有变化，再用少量50% 氢氧化钠溶液或浓盐酸调节，冰箱保存。

6、5柠檬酸柠檬酸钠缓冲液（0.1 mol/L）pH0.1   mol/L柠檬酸（毫升）0.1   mol/L柠檬酸钠（毫升）pH0.1   mol/L柠檬酸（毫升）0.1   mol/L柠檬酸钠（毫升）3.03.23.43.63.84.04.24.44.64.818.617.216.014.914.013.112.311.410.39.21.42.84.05.16.06.97.78.69.710.85.05.25.45.65.86.06.26.46.68.27.36.45.54.73.82.82.01.411.812

7、.713.614.515.316.217.218.018.6柠檬酸C6H8O7·H2O：分子量210.14，0.1 mol/L溶液为21.01克/升。柠檬酸钠Na3 C6H5O7·2H2O：分子量294.12，0.1 mol/L溶液为29.41克/毫升。 6乙酸乙酸钠缓冲液（0.2 mol/L） pH(180.2   mol/LNaAc(毫升0.3   mol/LHAc(毫升 pH(180.2   mol/LNaAc(毫升0.3   mol/LHAc(毫升2.63.

8、84.04.24.44.60.751.201.802.653.704.909.258.808.207.356.305.104.85.05.25.45.65.85.907.007.908.609.109.404.103.002.101.400.900.60Na2Ac·3H2O分子量 = 136.09，0.2 mol/L溶液为27.22克/升。 7磷酸盐缓冲液（1）磷酸氢二钠磷酸二氢钠缓冲液（0.2） pH0.2   mol/LNa2HPO4(毫升0.3   mol/LNaH2PO4(毫升 pH0.2 

9、60; mol/L Na2HPO4(毫升0.3   mol/LNaH2PO4(毫升5.85.96.06.16.26.36.46.56.66.76.86.98.010.012.315.018.522.526.531.537.543.549.555.092.090.087.785.081.577.573.568.562.556.551.045.07.07.17.27.37.47.57.67.77.87.98.061.067.072.077.081.084.087.089.591.593.094.739.033.028.023.019.016.013.010.58.57.05.3

10、Na2HPO4·2H2O分子量 = 178.05，0.2 mol/L溶液为85.61克/升。Na2HPO4·2H2O分子量 = 358.22，0.2 mol/L溶液为71.64克/升。Na2HPO4·2H2O分子量 = 156.03，0.2 mol/L溶液为31.21克/升。（2）磷酸氢二钠磷酸二氢钾缓冲液（1/15 mol/L）pHmol/L/15Na2HPO4(毫升mol/L/15KH2PO4(毫升pHmol/L/15Na2HPO4(毫升mol/L/15KH2PO4(毫升4.925.295.916.246.476.646.816.980.100.501.002

11、.003.004.005.006.009.909.509.008.007.006.005.004.007.177.387.738.048.348.678.187.008.009.009.509.759.9010.003.002.001.000.500.250.100Na2HPO4·2H2O分子量 = 178.05，1/15M溶液为11.876克/升。KH2PO4分子量 = 136.09，1/15M溶液为9.078克/升。 8磷酸二氢钾氢氧化钠缓冲液（0.05M）X毫升0.2M K2PO4 + Y毫升0.2N NaOH加水稀释至29毫升pH(20X (毫升Y（毫升）pH(20

12、X (毫升Y（毫升）5.86.06.26.46.66.85555550.3720.5700.8601.2601.7802.3657.07.27.47.67.88.05555552.9633.5003.9504.2804.5204.680 9巴比妥钠-盐酸缓冲液（18）pH0.04M巴比妥钠溶液0.2V盐酸pH0.04M巴比妥钠溶液(毫升0.2N盐酸(毫升6.87.07.27.47.67.88.08.210010010010010010010010018.417.816.715.313.411.479.397.218.48.68.89.09.29.49.6100100100100100

13、1001001005.213.822.521.651.130.700.35巴比妥钠盐分子量=206.18;0.04M溶液为8.25克/升 10Tris盐酸缓冲液（0.05M，25）50毫升0.1M三羟甲基氨基甲烷（Tris）溶液与X毫升0.1N盐酸混匀后，加水稀释至100毫升。pHX(毫升pHX(毫升7.107.207.307.407.507.607.707.807.908.0045.744.743.442.040.338.536.634.532.029.28.108.208.308.408.508.608.708.808.9026.222.919.917.214.712.410.3

14、8.57.0三羟甲基氨基甲烷（Tris）HOCH2 CH2OHCHOCH2 NH2分子量=121.14;1M溶液为12.114克/升。Tris溶液可从空气中吸收二氧化碳，使用时注意将瓶盖严。 11硼酸硼砂缓冲液（0.2M硼酸根）pH0.05M硼砂（毫升）0.2M硼砂（毫升）pH0.05M硼砂（毫升）0.2M硼酸（毫升）7.47.67.88.01.01.52.03.09.08.58.07.08.28.48.79.03.54.56.08.06.55.54.02.0硼砂Na2B4O7·H2O,分子量=381.43;0.05M溶液（=0.2M硼酸根）含19.07克/升。硼酸H2BO

15、3,分子量=61.84,0.2M溶液为12.37克/升。硼砂易失去结晶水，必须在带塞的瓶中保存。 11硼酸-硼砂缓冲液（0.2M硼酸根）pH0.05M硼砂（毫升）0.2M硼酸（毫升）pH0.05M硼砂（毫升）0.2M硼酸（毫升）7.47.67.88.01.01.52.03.09.08.58.07.08.28.48.79.03.54.56.08.06.55.54.02.0硼砂Na2B4O7·10H2O,分子量=381.43;0.05M溶液（=0.2M硼酸根）含19.07克/升。硼酸H2BO3，分子量=61.84, 0.2M溶液为12.37克/升。硼砂 易失去结晶水，必须在带塞

16、的瓶中保存。 12甘氨酸氢氧化钠缓冲液（0.05M）X毫升0.2M甘氨酸+Y毫升0.2NnaOH加水稀释至200毫升pHXYpHXY8.68.89.09.29.450505050504.06.08.812.016.89.69.810.010.410.6505050505022.427.232.038.645.5甘氨酸分子量=75.07;0.2M溶液含15.01克/升。 13硼砂-氢氧化钠缓冲液（0.05M硼酸根）X毫升0.05M硼砂+Y毫升0.2NNaOH加水稀释至200毫升pHXYpHXY9.39.49.65050506.011.023.09.810.010.150505

17、034.043.046.0硼砂Na2B4O7·10H2O,分子量=381.43;0.05M溶液为19.07克/升。 14碳酸钠-碳酸氢钠缓冲液（0.1M）Ca2+、Mg2+存在时不得使用pH0.1MNa2CO3(毫升0.1MN2HCO3(毫升20379.169.409.519.789.9010.1410.2810.5310.838.779.129.409.509.729.9010.0810.2810.57123456789987654321Na2CO2·10H2O分子量=286.2;0.1M溶液为28.62克/升。N2HCO3分子量=84.0;0.1M溶液为8.4

18、0克/升。 15“PBS”缓冲液PH7.67.47.27.0H2ONaClNa2HPO4NaH2PO410008.52.20.110008.52.20.210008.52.20.3100ml8.5g2.2g0.4g 三、实验室中常用酸碱的比重和浓度名称分子式分子量比重面分浓度%（W/W）当量浓度（粗略）N配1升1N溶液所需毫升数盐酸HCl36.471.1937.212.084   1.1835.411.8    1.1020.06.0 硫酸H2SO498.091.8495.636.028

19、0;  1.1824.86.0 硝酸HNO263.021.4270.9816.063   1.4065.314.5    1.2032.366.1 冰乙酸CH3COOH60.051.0599.517.459乙酸CH3COOH98.06 366.0 磷酸H2PO435.051.7185.015,30,45(依反应而定67(以15N计氨水NH4OH 0.90 1567   0.90427.014.370 

20、60; 0.9125.013.4    0.9610.05.6 氢氧化钠溶液NaOH40.01.550.01953 四、硫酸铵饱和度的常用表1调整硫酸铵溶液饱和度计算表（25）硫整硫酸铵终浓度,%饱和度  10202530333540455055606570758090100每1升溶液加固体硫酸铵的克数硫酸铵初浓度，%饱和度05611414417619620924327731335139043047251656166276710 578611813715018321625128832636540

21、644949459269420  2959789112315519022526230034038242452061925   3049619312515819323026730734839048558330    1930629412716219823527331435644954633     12437410714217721425229233342652235      3163941291

22、6420023817831941150640       31639713216820524528537546945        32659913417121025033943150         336610113717621430239255         &

23、#160;336710314117926435360           346910514322731465            347010719027570             357215323775  &

24、#160;           3611519880               7715790                79在25下，硫酸铵溶液由初浓度调到终浓度时，每升溶

25、液所加固体硫酸铵的克数。2 调整硫酸铵溶液饱和度计算表（0）在0下，硫酸铵溶液由初浓度调到终浓度时，每100毫升溶液所加固体硫酸铵的克数。 常见蛋白质分子量参考值 （单位：dalton）蛋白质分子量肌球蛋白myosin甲状腺球蛋白thyroglobulin-半乳糖苷酶-galactosidase副肌球蛋白paramyosin磷酸化酶aphosphorylase a血清白蛋白serum albuminL-氨基酸氧化酶L-amino acid oxidase地氧化氢酶catalase丙酮酸激活酶pyruvate kinase谷氨酸脱氢酶glutamate dehydrogenase亮氨酸

26、氨肽酶glutamae dehydrogenase-球蛋白，H链-globulin, H chain延胡索酸酶（反丁烯二酸酶）fumarase卵白蛋白ovalbumin醇脱氢酶（肝）alcohol dehydrogenase (liver烯醇酶enolase醛缩酶aldolase肌酸激酶creatine kinase胃蛋白酶原pepsinogenD-氨基酸氧化酶D-amino acid oxidase醇脱氢酶（酵母）alcohol dehydrogenase (yeast甘油醛磷酸脱氢酶dlyceraldehyde phosphate dehydrogenase原肌球蛋白tropomyosin

27、乳酸脱氢酶lactate dehydrgenase胃蛋白酶pepsin转磷酸核糖基酶phosphoribosyl transferase天冬氨酸氨甲酰转移酶，C链aspertate transcarbamylase, C chain羧肽酶 Acarboxypeptidase A碳酸酐酶carbonic anhydrase枯草杆菌蛋白酶subtilisin-球蛋白，L链-blobulin,L chain糜蛋白酶原（胰凝乳蛋白酶原）chymotrypsinogen胰蛋白酶trypsin木瓜蛋白酶（羧甲基）papain (carboxymethyl-乳球蛋白-lactoglobulin烟草花叶病毒外

28、壳蛋白（TWV外壳蛋白）TWV coat protein肌红蛋白myoglobin天门冬氨酸氨甲酰转移酶，R链aspartate transcarbamylase, R chain血红蛋白h(aemoglobinQ外壳蛋白Q coat protein溶菌酶lysozymeR17外壳蛋白R17 coat protein核糖核酸酶ribonuclease或RNase细胞色素Ccytochrome C糜蛋白酶（胰凝乳蛋白酶）chymotrypsin220,000165,000130,000100,00094,00068,00063,00060,00057,00053,00053,00050,0004

29、9,00043,00041,00041,00040,00040,00040,00037,00037,00036,00036,00036,00035,00035,00034,00034,00029,00027,60023,50025,70023,30023,00018,40017,50017,20017,00015,50015,00014,30013,75013,70011,70011,000或13,000 常见蛋白质等电点参考值 （单位：pH）蛋白质等电点鲑精蛋白salmine鲱精蛋白clupeine鲟精蛋白sturline胸腺组蛋白thymohistone珠蛋白（人）globin(

30、human卵白蛋白ovalbuin伴清蛋白conal bumin血清白蛋白serum albumin肌清蛋白myoal bumin肌浆蛋白myogen A-乳球蛋白-lactoglobulin卵黄蛋白livetin1球蛋白（人）1-globulin(human2球蛋白（人）2-globulin(human肌球蛋白Amyosin A原肌球蛋白myosin A铁传递蛋白siderophilin胎球蛋白fetuin血纤蛋白原fibrinogen-眼晶体蛋白-crystallin-眼晶体蛋白-crystallin花生球蛋白arachin伴花生球蛋白conarrachin角蛋白类keratins还原角蛋

31、白keratein胶原蛋白collagen鱼胶ichthyocol白明胶gelatin-酷蛋白-casein-酷蛋白-casein-酷蛋白-casein-卵清粘蛋白-ovomucoid1-粘蛋白1-mucoprotein卵黄类粘蛋白vitellomucoid尿促性腺激素urinary gonadotropin溶菌酶lyso zyme肌红蛋白myoglobin血红蛋白（人）hemoglobin(human血红蛋白（鸡）hemoglobin(hen血红蛋白（马）hemoglobin(horse血蓝蛋白hemerythrin蚯蚓血红蛋白chlorocruorin血绿蛋白chlorocruorin无脊

32、椎血红蛋白erythrocruorins细胞色素Ccytochrome C视紫质rhodopsin促凝血酶原激酶thromboplastin1-脂蛋白1-lipoprotein1-脂蛋白1-lipoprotein-卵黄脂磷蛋白-lipovitellin芜菁黄花病毒turnip yellow vvirus牛痘病毒vaccinia virus生长激素somatotropin催乳激素prolactin胰岛素insulin胃蛋白酶pepsin糜蛋白酶（胰凝乳蛋白酶chymotrypsin牛血清白蛋白bovine serum albumin核糖核酸酶（牛胰）ribonuclease或Rnase(bovi

33、ne pancreas甲状腺球蛋白thyroglobulin胸腺核组蛋白thymonucleohistone12.112.111.7110.87.54.71; 4.596.8;7.14.7-4.93.56.35.1-5.34.8-5.05.8;6.67.3;8.25.2-5.55.15.93.4-3.55.5-5.84.86.05.13.93.7-5.04.6-4.76.5-6.84.8-5.24.7-5.04.0-4.14.55.8-6.03.83-4.411.8-2.75.53.2-3.311.0-11.26.997.077.236.924.6-6.45.64.3-4.54.6-6.29.8-10.14