標(biāo)準(zhǔn)緩沖液的配制方法和PH計(jì)校正液(帶目錄).doc

標(biāo)準(zhǔn)緩沖液的配制及常用數(shù)據(jù)一、標(biāo)準(zhǔn)緩沖液pH值與溫度對(duì)照表2二、常用緩沖溶液的配制方法21 甘氨酸鹽酸緩沖液（0.05mol/L）22鄰苯二甲酸鹽酸緩沖液（0.05 mol/L）23磷酸氫二鈉檸檬酸緩沖液24檸檬酸氫氧化鈉-鹽酸緩沖液35檸檬酸檸檬酸鈉緩沖液（0.1 mol/L）36乙酸乙酸鈉緩沖液（0.2 mol/L）37磷酸鹽緩沖液38磷酸二氫鉀氫氧化鈉緩沖液（0.05M）49巴比妥鈉-鹽酸緩沖液（18）410Tris鹽酸緩沖液（0.05M，25）51TE Buffer550TAE Buffer 配制方法：5馬來酸 M=116511硼酸硼砂緩沖液（0.2M硼酸根）511硼酸-硼砂緩沖液（0.2M硼酸根）612甘氨酸氫氧化鈉緩沖液（0.05M）613硼砂-氫氧化鈉緩沖液（0.05M硼酸根）614碳酸鈉-碳酸氫鈉緩沖液（0.1M）615“PBS”緩沖液7三、實(shí)驗(yàn)室中常用酸堿的比重和濃度7四、硫酸銨飽和度的常用表8常見蛋白質(zhì)分子量參考值 （單位：dalton）8常見蛋白質(zhì)等電點(diǎn)參考值 （單位：pH）9元素的原子量表11PH計(jì)校正溶液配置的標(biāo)準(zhǔn)方法13一、標(biāo)準(zhǔn)緩沖液pH值與溫度對(duì)照表溫度四草酸氫鉀0.05M鄰苯二甲酸氫鉀0.05M混合磷酸鹽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、常用緩沖溶液的配制方法1 甘氨酸鹽酸緩沖液（0.05mol/L）X毫升0.2 mol/L甘氨酸+Y毫升0.2 mol/L HCI，再加水稀釋至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 = 75.07，0.2 mol/L甘氨酸溶液含15.01克/升。2鄰苯二甲酸鹽酸緩沖液（0.05 mol/L）X毫升0.2 mol/L鄰苯二甲酸氫鉀 + 0.2 mol/L HCl，再加水稀釋到20毫升pH(20)XYpH(20)XY2.22.42.62.83.0555554.0703.9603.2952.6422.0223.23.43.63.855551.4700.9900.5970.263鄰苯二甲酸氫鉀分子量 = 204.23，0.2 mol/L鄰苯二甲酸氫溶液含40.85克/升3磷酸氫二鈉檸檬酸緩沖液pH0.2mol/L Na2HPO4（毫升）0.1mol/L檸檬酸（毫升）pH0.2mol/L Na2HPO4 （毫升）0.1mol/L檸檬酸（毫升）2.22.42.62.83.03.23.43.63.84.04.24.44.64.85.00.401.242.183.174.114.945.706.447.107.718.288.829.359.8610.3010.6018.7617.8216.8315.8915.0614.3013.5612.9012.2911.7211.1810.6510.149.705.25.45.65.86.06.26.46.66.87.07.27.47.67.88.010.7211.1511.6012.0912.6313.2213.8514.5515.4516.4717.3918.1718.7319.1519.459.288.858.407.917.376.786.155.454.553.532.611.831.270.850.55Na2HPO4分子量 = 14.98，0.2 mol/L溶液為28.40克/升。Na2HPO4-2H2O分子量 = 178.05，0.2 mol/L溶液含35.01克/升。C4H2O7H2O分子量 = 210.14，0.1 mol/L溶液為21.01克/升。4檸檬酸氫氧化鈉-鹽酸緩沖液pH鈉離子濃度（mol/L）檸檬酸（克）C6H8O7H2O氫氧化鈉(克)NaOH 97%鹽酸（毫升）HCl（濃）最終體積（升）2.23.13.34.35.35.86.50.200.200.200.200.350.450.3821021021021024528526684838383144186156160116106456810512610101010101010 使用時(shí)可以每升中加入1克克酚，若最后pH值有變化，再用少量50%氫氧化鈉溶液或濃鹽酸調(diào)節(jié)，冰箱保存。5檸檬酸檸檬酸鈉緩沖液（0.1 mol/L）pH0.1 mol/L檸檬酸（毫升）0.1 mol/L檸檬酸鈉（毫升）pH0.1 mol/L檸檬酸（毫升）0.1 mol/L檸檬酸鈉（毫升）3.03.23.43.63.84.04.24.44.64.818.617.216.014.914.013.112.311.410.39.21.42.84.05.16.06.97.78.69.710.85.05.25.45.65.86.06.26.46.68.27.36.45.54.73.82.82.01.411.812.713.614.515.316.217.218.018.6檸檬酸C6H8O7H2O：分子量210.14，0.1 mol/L溶液為21.01克/升。檸檬酸鈉Na3 C6H5O72H2O：分子量294.12，0.1 mol/L溶液為29.41克/毫升。6乙酸乙酸鈉緩沖液（0.2 mol/L）pH(18)0.2 mol/LNaAc(毫升)0.3 mol/L HAc(毫升)pH(18)0.2 mol/LNaAc(毫升)0.3 mol/LHAc(毫升)2.63.84.04.24.44.60.751.201.802.653.704.909.258.808.207.356.305.104.85.05.25.45.65.85.907.007.908.609.109.404.103.002.101.400.900.60Na2Ac3H2O分子量 = 136.09，0.2 mol/L溶液為27.22克/升。7磷酸鹽緩沖液（1）磷酸氫二鈉磷酸二氫鈉緩沖液（0.2）pH0.2 mol/L Na2HPO4(毫升)0.3 mol/L NaH2PO4(毫升)pH0.2 mol/L Na2HPO4(毫升)0.3 mol/L NaH2PO4(毫升)5.85.96.06.16.26.36.46.56.66.76.86.98.010.012.315.018.522.526.531.537.543.549.555.092.090.087.785.081.577.573.568.562.5 56.551.045.07.07.17.27.37.47.57.67.77.87.98.061.067.072.077.081.084.087.089.591.593.094.739.033.028.023.019.016.013.010.58.57.05.3Na2HPO42H2O分子量 = 178.05，0.2 mol/L溶液為85.61克/升。Na2HPO42H2O分子量 = 358.22，0.2 mol/L溶液為71.64克/升。Na2HPO42H2O分子量 = 156.03，0.2 mol/L溶液為31.21克/升。（2）磷酸氫二鈉磷酸二氫鉀緩沖液（1/15 mol/L）pHmol/L/15Na2HPO4(毫升)mol/L/15KH2PO4(毫升)pHmol/L/15Na2HPO4(毫升)mol/L/15KH2PO4(毫升)4.925.295.916.246.476.646.816.980.100.501.002.003.004.005.006.009.909.509.008.007.006.005.004.007.177.387.738.048.348.678.187.008.009.009.509.759.9010.003.002.001.000.500.250.100Na2HPO42H2O分子量 = 178.05，1/15M溶液為11.876克/升。KH2PO4分子量 = 136.09，1/15M溶液為9.078克/升。8磷酸二氫鉀氫氧化鈉緩沖液（0.05M）X毫升0.2M K2PO4 + Y毫升0.2N NaOH加水稀釋至29毫升 pH(20)X (毫升)Y（毫升）pH(20)X (毫升)Y（毫升）5.86.06.26.46.66.85555550.3720.5700.8601.2601.7802.3657.07.27.47.67.88.05555552.9633.5003.9504.2804.5204.6809巴比妥鈉-鹽酸緩沖液（18）pH0.04M巴比妥鈉溶液0.2V鹽酸pH0.04M巴比妥鈉溶液(毫升)0.2N鹽酸(毫升)6.87.07.27.47.67.88.08.210010010010010010010010018.417.816.715.313.411.479.397.218.48.68.89.09.29.49.61001001001001001001001005.213.822.521.651.130.700.35巴比妥鈉鹽分子量=206.18;0.04M溶液為8.25克/升10Tris鹽酸緩沖液（0.05M，25）50毫升0.1M三羥甲基氨基甲烷（Tris）溶液與X毫升0.1N鹽酸混勻后，加水稀釋至100毫升。pHX(毫升)pHX(毫升)7.107.207.307.407.507.607.707.807.908.0045.744.743.442.040.338.536.634.532.029.28.108.208.308.408.508.608.708.808.9026.222.919.917.214.712.410.38.57.0三羥甲基氨基甲烷（Tris）HOCH2 CH2OH CHOCH2 NH2分子量=121.14;1M溶液為12.114克/升。Tris溶液可從空氣中吸收二氧化碳，使用時(shí)注意將瓶蓋嚴(yán)。1TE Buffer組成濃度：10mM Tris-HCl 1mM EDTA PH=8.0 配制量：500ml 配制方法：量取下列溶液于500ml燒杯中 1M Tris-HCl Buffer PH=8.0 5ml 0.5M EDTA PH=8.0 1ml 向燒杯中加入約400ml dd H2O均勻混合；將溶液定容到500ml后，高溫高壓滅菌；室溫保存.50TAE Buffer 配制方法： 1。稱量Tris 242g，Na2EDTA2H2O 37.2g于1L燒杯中； 2。向燒杯中加入約800ml去離子水，充分?jǐn)嚢杈鶆颍?3。加入57.1ml的冰乙酸，充分溶解； 4。用NaOH調(diào)pH至8.3，加去離子水定容至1L后，室溫保存。 使用時(shí)稀釋50倍（取20ml稀釋至1L）或100倍 即1TAE Buffer 或 0.5TAE馬來酸 M=11611硼酸硼砂緩沖液（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Na2B4O7H2O,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溶液為12.37克/升。硼砂易失去結(jié)晶水，必須在帶塞的瓶中保存。11硼酸-硼砂緩沖液（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Na2B4O710H2O,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溶液為12.37克/升。硼砂易失去結(jié)晶水，必須在帶塞的瓶中保存。12甘氨酸氫氧化鈉緩沖液（0.05M）X毫升0.2M甘氨酸+Y毫升0.2NnaOH加水稀釋至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-氫氧化鈉緩沖液（0.05M硼酸根）X毫升0.05M硼砂+Y毫升0.2NNaOH加水稀釋至200毫升pHXYpHXY9.39.49.65050506.011.023.09.810.010.150505034.043.046.0硼砂Na2B4O710H2O,分子量=381.43;0.05M溶液為19.07克/升。14碳酸鈉-碳酸氫鈉緩沖液（0.1M）Ca2+、Mg2+存在時(shí)不得使用pH0.1MNa2CO3(毫升)0.1MN2HCO3(毫升)20379.169.409.519.789.9010.1410.2810.5310.838.779.129.409.509.729.9010.0810.2810.57123456789987654321Na2CO210H2O分子量=286.2;0.1M溶液為28.62克/升。N2HCO3分子量=84.0;0.1M溶液為8.40克/升。15“PBS”緩沖液PH7.67.47.27.0H2ONaClNa2HPO4NaH2PO410008.52.20.110008.52.20.210008.52.20.3100ml8.5g2.2g0.4g三、實(shí)驗(yàn)室中常用酸堿的比重和濃度名稱分子式分子量比重面分濃度%（W/W）當(dāng)量濃度（粗略）N配1升1N溶液所需毫升數(shù)鹽酸HCl36.471.1937.212.0841.1835.411.81.1020.06.0硫酸H2SO498.091.8495.636.0281.1824.86.0硝酸HNO263.021.4270.9816.0631.4065.314.51.2032.366.1冰乙酸CH3COOH60.051.0599.517.459乙酸CH3COOH98.06366.0磷酸H2PO435.051.7185.015,30,45(依反應(yīng)而定)67(以15N計(jì))氨水NH4OH0.9015670.90427.014.3700.9125.013.40.9610.05.6氫氧化鈉溶液NaOH40.01.550.01953四、硫酸銨飽和度的常用表1調(diào)整硫酸銨溶液飽和度計(jì)算表（25）硫整硫酸銨終濃度,%飽和度10202530333540455055606570758090100每1升溶液加固體硫酸銨的克數(shù)硫酸銨初濃度，%飽和度05611414417619620924327731335139043047251656166276710578611813715018321625128832636540644949459269420295978911231551902252623003403824245206192530496193125158193230267307348390485583301930629412716219823527331435644954633124374107142177214252292333426522353163941291642002381783194115064031639713216820524528537546945326599134171210250339431503366101137176214302392553367103141179264353603469105143227314653470107190275703572153237753611519880771579079在25下，硫酸銨溶液由初濃度調(diào)到終濃度時(shí)，每升溶液所加固體硫酸銨的克數(shù)。2 調(diào)整硫酸銨溶液飽和度計(jì)算表（0）在0下，硫酸銨溶液由初濃度調(diào)到終濃度時(shí)，每100毫升溶液所加固體硫酸銨的克數(shù)。常見蛋白質(zhì)分子量參考值 （單位：dalton）蛋白質(zhì)分子量肌球蛋白myosin甲狀腺球蛋白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地氧化氫酶catalase丙酮酸激活酶pyruvate kinase谷氨酸脫氫酶glutamate dehydrogenase亮氨酸氨肽酶glutamae dehydrogenase-球蛋白，H鏈-globulin, H chain延胡索酸酶（反丁烯二酸酶）fumarase卵白蛋白o(hù)valbumin醇脫氫酶（肝）alcohol dehydrogenase (liver)烯醇酶enolase醛縮酶aldolase肌酸激酶creatine kinase胃蛋白酶原pepsinogenD-氨基酸氧化酶D-amino acid oxidase醇脫氫酶（酵母）alcohol dehydrogenase (yeast)甘油醛磷酸脫氫酶dlyceraldehyde phosphate dehydrogenase原肌球蛋白tropomyosin乳酸脫氫酶lactate dehydrgenase胃蛋白酶pepsin轉(zhuǎn)磷酸核糖基酶phosphoribosyl transferase天冬氨酸氨甲酰轉(zhuǎn)移酶，C鏈aspertate transcarbamylase, C chain羧肽酶 Acarboxypeptidase A碳酸酐酶carbonic anhydrase枯草桿菌蛋白酶subtilisin-球蛋白，L鏈-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煙草花葉病毒外殼蛋白（TWV外殼蛋白）TWV coat protein肌紅蛋白myoglobin天門冬氨酸氨甲酰轉(zhuǎn)移酶，R鏈aspartate transcarbamylase, R chain血紅蛋白h(a)emoglobinQ外殼蛋白Q coat protein溶菌酶lysozymeR17外殼蛋白R(shí)17 coat protein核糖核酸酶ribonuclease或RNase細(xì)胞色素Ccytochrome C糜蛋白酶（胰凝乳蛋白酶）chymotrypsin220,000165,000130,000100,00094,00068,00063,00060,00057,00053,00053,00050,00049,00043,00041,00041,00040,00040,00040,00037,00037,00036,00036,00036,00035,00035,00034,00034,00029,00027,60023,50025,70023,30023,00018,40017,50017,20017,00015,50015,00014,30013,75013,70011,70011,000或13,000常見蛋白質(zhì)等電點(diǎn)參考值 （單位：pH）蛋白質(zhì)等電點(diǎn)鮭精蛋白salmine鯡精蛋白clupeine鱘精蛋白sturline胸腺組蛋白thymohistone珠蛋白（人）globin(human)卵白蛋白o(hù)valbuin伴清蛋白conal bumin血清白蛋白serum albumin肌清蛋白myoal bumin肌漿蛋白myogen A-乳球蛋白-lactoglobulin卵黃蛋白livetin1球蛋白（人）1-globulin(human)2球蛋白（人）2-globulin(human)肌球蛋白Amyosin A原肌球蛋白myosin A鐵傳遞蛋白siderophilin胎球蛋白fetuin血纖蛋白原fibrinogen-眼晶體蛋白-crystallin-眼晶體蛋白-crystallin花生球蛋白arachin伴花生球蛋白conarrachin角蛋白類keratins還原角蛋白keratein膠原蛋白collagen魚膠ichthyocol白明膠gelatin-酷蛋白-casein-酷蛋白-casein-酷蛋白-casein-卵清粘蛋白-ovomucoid1-粘蛋白1-mucoprotein卵黃類粘蛋白vitellomucoid尿促性腺激素urinary gonadotropin溶菌酶lyso zyme肌紅蛋白myoglobin血紅蛋白（人）hemoglobin(human)血紅蛋白（雞）hemoglobin(hen)血紅蛋白（馬）hemoglobin(horse)血藍(lán)蛋白hemerythrin蚯蚓血紅蛋白chlorocruorin血綠蛋白chlorocruorin無脊椎血紅蛋白erythrocruorins細(xì)胞色素Ccytochrome C視紫質(zhì)rhodopsin促凝血酶原激酶thromboplastin1-脂蛋白1-lipoprotein1-脂蛋白1-lipoprotein-卵黃脂磷蛋白-lipovitellin蕪菁黃花病毒turnip yellow vvirus牛痘病毒vaccinia virus生長(zhǎng)激素somatotropin催乳激素prolactin胰島素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(bovine pancreas)甲狀腺球蛋白thyroglobulin胸腺核組蛋白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;8.25.2-5.55.15.93.4-3.55.5-5.84.86.05.13.93.7-5.04.6-4.76.5-6.84.8-5.24.7-5.04.0-4.14.55.8-6.03.83-4.411.8-2.75.53.2-3.311.0-11.26.997.077.236.924.6-6.45.64.3-4.54.6-6.29.8-10.14.47-4.575.25.55.45.93.755.36.855.735.351.0左右8.14.97.84.584左右元素的原子量表（錄自1997年國際原子量表，并全部取四位有效數(shù)字）元素符號(hào)原子量原子序數(shù)錒銀鋁镅氬砷砹金硼鋇鈹鉍锫溴碳鈣鎘鈰锎氯鋦鈷鉻銫銅鏑鑷锿銪氟鐵鐨鈁鎵釓鍺氫氦鉿汞鈥碘銦銥鉀氪鑭鋰镥鐒鍆鎂錳鉬氮鈉鈮釹氖鎳锘镎氧鋨磷鎂鉛鈀钷釙鐠鉑钚鐳銣錸銠氡釕硫銻鈧硒硅釤錫鍶鉭鋱锝碲釷鈦鉈銩鈾釩鎢氙釔鐿鋅鋯AcAgAlAmArAsAtAuBBaBeBiBkBrCCaCdCeCfClCmCoCrCsCuDyErEsEuFFeFmFrGaGdGeHHeHfHgHoIInIrKKrLaLiLuLrMdMgMnMoNNaNbNdNeNiNoNpOOSPPaPbPdPmPoPrP