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1、PBS緩沖液配方不管是免疫組化,還是細(xì)胞培養(yǎng)中,常用到PBS緩沖液,下面是我們實(shí)驗(yàn)室的配方,供大家參考。我們實(shí)驗(yàn)室的配方:A液:0.1mol/L磷酸二氫鉀:磷酸二氫鉀(KH2PO4,MW136.09)1.361g,加雙蒸水至100ml。B液:0.1mol/L磷酸氫二鈉:酸酸氫二鈉(Na2HPO4.2H2O,MW177.99)1.78g,加雙蒸水至100ml?;蛘咚崴釟涠c(Na2HPO4.12H2O,MW358.14)3.58g,加雙蒸水至100ml。pH--A液(ml)--B液(ml)5.60--9.50--0.255.91--9.00--1.
2、006.24--8.00--2.006.47--7.00--3.006.64--6.00--4.006.81--5.00--5.006.98--4.00--6.007.17--3.00--7.007.38--2.00--8.007.73--1.00--9.008.04--0.50--9.50一般免疫組化用PBS都只包含Na2HPO4?12H2O(磷酸氫二鈉)NaH2PO4?2H2O(磷酸二氫鈉)Nacl用于細(xì)胞培養(yǎng)的PBS需含有氯化鉀稱取8gNaCl、0.2gKCl、1.44gNa2HPO4和0.24gKH2PO4,溶于800ml蒸餾水中,用HC
3、l調(diào)節(jié)溶液的pH值至7.4,最后加蒸餾水定容至1L即可0.01MPBS緩沖液稱取NaCl?8g,KCl?0.2g,Na2HPO4?12H2O?3.63g,?KH2PO4?0.24g,溶于900ml雙蒸水中,用鹽酸調(diào)pH值至7.4,加水定容至1L,常溫保存?zhèn)溆?。PBS是最普遍不過的實(shí)驗(yàn)室緩沖液,但其配方各異。以下是我的總結(jié):?母液的配制:?0.2M?Na2HPO4:稱取?71.6g?Na2HPO4-12H2O,溶于?1000ml?水0.2M?NaH2PO4:稱取?31.2g?NaH2PO4-2H2O,溶于1000ml?水各種濃度PB(pH=7.4)
4、的配制:先配?0.2M?PB?(pH=7.4,100ml):取19ml?0.2mol/L的?NaH2PO4,?81ml?0.2mol/L?的?Na2HPO4,?即可。然后?只需將0.2M?PB?(pH=7.4)按相應(yīng)比例適當(dāng)稀釋?即可,如:0.1M?PB(PH=7.4)?:取?500ml?0.2M?PB,加水稀釋至?1000ml?即可。0.01M?PB?(PH=7.4):取50ml?0.2M?PB,加水稀釋至?1000ml?即可。0.02M?PB?(PH=7.4):取100ml?0.2?M?PB,加水稀釋至?1000ml?即可。?若需要?NaCL
5、?的話,加入?NaCL?至0.9%(g/100ml)即可。?另:其它各種另?PH值的?0.2M?PB(100ml)配方?:pH???????0.2M?NaH2PO4(ml)???0.2M?Na2HPO4(ml)5.7?????????????93.5??????????????????????????6.55.8?????????????92?????????????????????????????8????????????5.9?????????????90?????????????????????????????106.0??????????
6、???87.7??????????????????????????12.36.1?????????????85?????????????????????????????156.2?????????????81.5??????????????????????????18.56.3?????????????77.5??????????????????????????22.56.4?????????????73.5??????????????????????????26.56.5?????????????68.5????????????????????
7、??????31.56.6?????????????62.5??????????????????????????37.56.7?????????????56.5??????????????????????????43.56.8?????????????51????????????????????????????496.9??????????????45???????????????????????????557.0??????????????38???????????????????????????627.1??????????????33???
8、????????????????????????677.2?????????????28????????????????????????