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PBS的配制

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PBS标准配方如下(1L): 成分 分子量 终浓度(mM) 用量(g) 近似用量(g) 半数用量(g) NaCl 58.44 137 8.006 8.00 4.00 KCl 74.551 2.7 0.201 0.20 0.10 Na2HPO4·12H2O 358.14 10 3.581 3.58 1.79 KH2PO4 136.086 1.76 0.240 0.24 0.12

理论上需要调pH,但在实际操作中,用蒸馏水或者超纯水配制后,pH在7.3左右,所以通常不用调pH。 10×PBS配方如下(1L): 成分 近似用量(g) NaCl 80.0 KCl 2.0 Na2HPO4·12H2O 35.8 KH2PO4 2.4

10×PBS保存于4℃时,会析出晶体,且不易复溶,因此10×PBS要在室温保存。 PBS在室温保存时容易长菌,应保存于4℃。

关于PBS配方的比较与讨论:

我们平时会用到PBS,但网上PBS的配方有多种,不知如何选择?

参考一:

看你的用途了。配方虽然有多种,但在“等渗”这个概念上相差都不大。常见的配方一般有三种: 1、0.9%的生理盐水

有些人也把它叫做PBS,实际上是错误的概念,它并没有“PB”,没有缓冲能力,纯化少用。 2、pH7.3,137mM NaCl,2.7mM KCl,10mM Na2HPO4,1.8mM KH2PO4。

经验证,这种配方是没有办法加钙离子和镁离子的,会形成沉淀。此配方为0.01M的PBS,所对应的质量分别为8g NaCl,0.2g KCl,1.44g(1.42g)Na2HPO4和0.24g(0.27g)KH2PO4。考虑到Na2HPO4通常为Na2HPO4·12H20,所以应该加3.58g。(修正过的)。

这个配方比较常用的,含K,特别在分子生物学上常用。通常所说的浓度0.01M指的是缓冲溶液中所有的磷酸根浓度,而非Na离子或K离子的浓度,Na离子和K离子只是用来调节渗透压的。如果是用于免疫组化的话,则需要在配置的时候分别加入100u/ml青霉素和链霉素之后再调pH、定容、灭菌消毒。 3、pH7.2~7.4,20mM PB,150mM NaCl

这个配方是蛋白质纯化常用的PBS,配制起来比较方便。

具体到称量方面,要注意试剂是否有结晶水,分子量是不同的。 参考二:

关键看用途,如果是要活性,要考虑你的目的蛋白在什么样的环境下稳定,主要是掌握好钠钾平衡和生理状态下离子强度,在此前提下有一定的缓冲能力。 参考三:

PBS是phosphate-buffered saline的意思。 看看下面这个,或许对你有用 Applications

PBS has many uses because it is isotonic and non-toxic to cells. It can be used to dilute substances. It is used to rinse containers containing cells. PBS can be used as a diluent in methods to dry biomolecules, as water molecules within it will be structured around the substance (protein, for example) to be 'dried' and immobilized to a solid surface. The thin film of water that binds to the substance prevents denaturation or other conformational changes. Carbonate buffers may be used for the same purpose but with less effectiveness. PBS can be used to take a reference spectrum when measuring the protein adsorption in ellipsometry.

Additives can be used to add function. For example, PBS with EDTA is also used to disengage attached and clumped cells. Divalent metals such as zinc, however, cannot be added as this will result in precipitation. For these types of applications, Good's buffers are recommended. Preparation

There are many different ways to prepare PBS. Some formulations do not contain potassium, while others contain calcium or magnesium. One of the most common preparations is described below.

The simplest way to prepare a PBS solution is to use PBS buffer tablets. They are formulated to give a ready to use PBS solution upon dissolution in a specified quantity of distilled water. They are available in the standard volumes: 100, 200, 500 and 1000 ml.

A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na2HPO4 and 24 g KH2PO4 in 8 L of distilled water, and topping up to 10 L. The pH is ~6.8, but when diluted to 1x PBS it should change to 7.4. When making buffer solutions, it is good practice to always measure the pH directly using a pH meter. If necessary, pH can be adjusted using hydrochloric acid or sodium hydroxide. On dilution, the resultant 1x PBS should have a final concentration of 137 mM NaCl, 2.7 mM KCl, 10 mM Sodium Phosphate dibasic, 2 mM

Potassium Phosphate monobasic and a pH of 7.4.

Another preparation is described in Molecular Cloning by Sambrook, Fritsch and Maniatis, Apendix B.12 as follows:

For 1 litre of 1x Phosphate-buffered saline (1x PBS buffer) use: - Dissolve in 800 ml of distilled H2O: - 8 g of NaCl (137mM) - 0.2 g of KCl (2.7mM)

- 1.44 g of Na2HPO4 (10mM) - 0.24 g of KH2PO4 (1.76mM)

- Adjust the pH to 7.4 with HCl or NaOH - Add H2O to 1 liter.

Dispense the solution into aliquots and sterilize them by autoclaving (20 min, 121℃, liquid cycle). Store at room temperature. 参考四:

含钾盐的PBS应该和生理条件最为接近,可能是保持某些生物大分子功能性的要求,如酶的活性。而在蛋白质纯化过程中,钠盐或钾盐所提供的就是离子强度,从方便性考虑,用一种就够了。钠盐最普通,就选择氯化钠了。但也不是绝对的,在GST亲和柱纯化中所用的平衡缓冲液就是含钾盐的PBS,可能因为它的结合是酶和底物之间的关系吧。

PBS(Phosphate-buffered Saline) 分子克隆上的配方 NaCl 8g 137mM KCl 0.2g 2.7mM Na2HPO4 1.44g 10mM KH2PO4 0.24g 2mM

溶到800ml蒸馏水中,用HCl调pH到7.4,然后加水补充到1000ml,15psi(1.034×105)20min高压灭菌或者滤膜除菌,室温保存。一般用于实验室中的PBS缓冲液都是这样配制,而不用摩尔分数表示。分子克隆上补充“PBS是一种通用试剂。值得指出的是本文列出的配方缺乏双价阳离子,如果需要,PBS可以补加成1mM CaCl2和0.5mM MgCl2”。但本人在试验过,加入后立马形成沉淀,不明何故。

免疫学手册上的配方: NaCl 19.00g 154mM Na2HPO4 1.15 g 8.1mM NaH2PO4 0.23 g 1.9mM

加水900 ml 调pH 7.2~7.4 再加水补充到1L

0.05M储存液 NaCl 100.00g Na2HPO4?12H2O 66.15g NaH2PO4?H2O 6.66g KCl 4.50g

加双蒸水至4500ml,用HCl或NaOH调pH到7.4即可。使用时稀释5倍,即0.01M。

PBS(0.1M,pH 7.4)贮存液配方(10倍浓缩) NaCl 80g Na2HPO4?12H2O 32.3g NaH2PO4?2H2O 4.5g 加蒸馏水至 1000ml 或者 NaCl 80g Na2HPO4?2H2O 11.5g KH2PO4 2g KCl 2g 加蒸馏水至 1000ml

天津百浩生物科技有限公司 PBS(0.01M) NaCl 8g Na2HPO4·12H2O 2.9g NaH2PO4·2H2O 0.2964g 加蒸馏水至 1000ml

takara公司:与分子克隆基本相同,不同之处为Na2HPO4 1.42g(分子克隆是1.44g),并且也提示可以加入钙镁离子。

PBS的配制

PBS标准配方如下(1L):成分分子量终浓度(mM)用量(g)近似用量(g)半数用量(g)NaCl58.441378.0068.004.00KCl74.5512.70.2010.200.10Na2HPO4·12H2O358.14103.5813.581.79
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