主题:【讨论】关于三种磷酸缓冲溶液的区别和配制方法

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关于磷酸缓冲溶液的知识,我在网上搜索了好多帖子,但是还没有满意的答案。现在把他们链接过来,希望有高手们能给出自己的见解。

PBS可以为三种溶液的英文缩写,分别是
1.磷酸盐缓冲溶液(phosphate buffered solution)、
2.磷酸盐缓冲盐水(phosphate buffered saline)及
3.磷酸盐缓冲钠(phosphate buffered sodium),
其配制方法不同,pH值不同,发挥的生物学作用亦不完全相同。
那么他们的区别是什么?各自的配制方法是怎么样的?



无钙镁离子磷酸缓冲盐水(PBS)
(0.15Mol/L PH 7.2)
NaCl 8.0g
KCl 0.2g
Na2HPO4 1.15g
(如为Na2HPO4·12H2O,则为2.89g)
KH2PO4 0.2g
将上述试剂依次溶于1 000ml去离子水中,完全溶解后,115℃高压灭菌10min~15min,存在4℃冰箱中备用。此液可用于配制和稀释细胞分散剂以及洗涤细胞培养物。


我们用的都是第二种
磷酸盐缓冲盐水(phosphate buffered sodium),为什么用第三种或第一种呢
lanping 2007-9-17 17:21
这么多年以来我都以为PBS就是磷酸盐缓冲溶液,其他两种说法的英语名称常见到,不过我一直都以为他们是差不多的东西。不同的汉语翻译倒是第一次听说。
我们公司用的PBS有很多种,抗体组用的中和PBS配方就比较简单,没有K+离子, 只起到缓冲PH的作用;Na2HPO4+ KH2PO4 按照不同比例混匀,可以得到不同PH的缓冲液,用于不同目的,这个在分子生物学中用得很多; 用于活细胞或者细菌处理的,为了保持渗透压,会加入其他的溶液,最常见的就是NaCl 和 KCl调节离子浓度。
以上是个人的理解,欢迎批评。
xydqq 2007-9-18 09:32
既然抗体组的PBS是起到缓冲PH的作用,那加入80G的NACL作用是什么?需要平衡渗透压吗?
还有加入K离子的是不是因为细胞上NA-K泵的原因啊
网络伯牙 2007-9-18 17:16
血清里面又没有细胞,何来NA-K泵一说啊? [s:49]
网络伯牙 2007-9-18 17:17
我觉得之所以这么配,其实就是对血清中缓冲环境的一种模拟罢了
xydqq 2007-9-18 17:23
引用第3楼网络伯牙于2007-09-18 17:16发表的:
血清里面又没有细胞,何来NA-K泵一说啊? [s:49]



我没说血清撒。。。。。。 我们用的又没加K,我是说加了K的的
xydqq 2007-9-18 17:24
不加NACL会怎么样啊?多省钱啊
lanping 2007-9-20 21:14
在中和磷酸盐缓冲液中,NaCl的主要作用应当是保持一定的离子浓度,以维持抗体的活性。在离子浓度过高或过低的情况下,蛋白质都会变性,抗体当然不例外。
xydqq 2007-9-21 00:19
哦 晓得了,0.8%的NACL差不多就是生理盐水的浓度。。。。。 [s:46]

ps:哺乳类需用生理盐水浓度是0.9%。称取0.9克氯化钠,溶解在少量蒸馏水中,稀释到100毫升。
鸟类需用的生理盐水浓度是0.75%。称取0.75克氯化钠,溶解后用蒸馏水稀释到100毫升。
两栖类需用的生理盐水浓度是0.65%。称取0.65克氯化钠,溶解后用蒸馏水稀释到100毫升。
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Phosphate buffered saline
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Phosphate buffered saline (abbreviated PBS) is a buffer solution commonly used in biochemistry. It is a salty solution containing sodium chloride, sodium phosphate and potassium phosphate. The buffer helps to maintain a constant pH. The concentration usually matches the human body (isotonic).

Applications
PBS has many uses because it is isotonic and non-toxic to cells. It can be used to dilute substances. It is used as a cellular cleaning solution. To ensure the prolonged dry-storage of immobilized-biomolecules like proteins, enzymatic proteins etc., PBS is used as biomolecule diluent since it can structure water around biomolecules immobilized to the solid surface. Such thin film of water prevents denaturing of biomolecules or conformational changes to them. Carbonate buffers may be used for the same purpose but with less effectiveness. PBS can be used as a reference spectra 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 cause precipitation. For these types of applications, Good's buffers are recommended.

Preparation
A 10 liter stock of 10x PBS can be prepared by dissolving 800 g NaCl, 20 g KCl, 144 g Na2HPO4 and 24g 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.

On dilution, the resultant 1x PBS will have a final concentration of 137 mM NaCl, 10 mM Phosphate, 2.7 mM KCl, pH 7.4
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Product Description
Name: Phosphate Buffered Saline Buffer
Chemical description: When reconstituded with water, PBS1x final concentrations are 137mM Sodium Chloride, 10mM phosphate, 2.7mM Potassium Chloride, pH is 7.4

Cat.number: UP68723A, 10L inquire for other quantities

Form: White powder, pack qsp 10L
Use: reconstitute PBS 1x with distilled water:
1 pack (9.88g), qsp 10L of d.water
mix; the solution should be clear and
without particulates
rem: One also may use 1pack with 1L to
prepare 10X PBS mother solution

Storage: Room temperature (.)

Applications: PBS is used in various laboratory technics, including immunodetection,biochemistry, purification, cell culture (ask for our Sterile PBS)… Phosphate buffer should not be used in assays where competition for phosphate groups, or complex formation
with a metal ion is essential for the enzyme activation. Phosphate ions will inhibit carboxypeptidase,carboxylase, urease, muscle diaminase, formase and phosphoglucomutase.

For any question, please contact Uptima, or our local distributor.
Hotline : +33(0)4 70 03 76 06; uptima@interchim.com ; http://www.interchim.com
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