
Application | Each vial is sufficient for a volume of 100ml solution |
---|---|
Just add to your cell lysis buffer |
The Xpert Phosphatase Inhibitor Cocktail Solution(100X)
is a mixture of several inhibitors to prevent valuable
proteinsfrom dephospholylation. Phosphorylation and
dephosphorylation is a molecular on/off switch that regulates
a number of key biological pathways within the cell,
including signal transduction, cell division, and apoptosis.
The Xpert Phosphatase Inhibitor Cocktail preserves
phosphorylation of proteins in cell lysates and tissue
extracts.The cocktail contains a mixture of four
phosphatase inhibitors of broad specificity, including
sodium fluoride, sodium orthovanadate, sodium
pyrophosphateand sodium glycerophosphate.
Unlike other commerciallyavailable phosphatase
inhibitor cocktails which protectagainst either
Ser/Thr phosphatases or protein tyrosine
phosphatases.
The Xpert Phosphatase Inhibitor Cocktail protects
phosphoproteins from both families of phosphatases.
Conveniently packaged in a 100X format, the cocktail
is stable at 4°C and is compatible with common protein
assays, including the BCA Protein Assay and Bradford-
basedassays such as Coomassie.
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TBST中含有Tris-Hcl,NaCl,Tween20这三种物质,是做WESTERNBLOT中常用的一种缓冲液。
TBST缓冲液的配制
1000ml×TBST的配置
先称量NaCl40g,倒入烧杯中,加DDW蒸馏水400ml,再称量NaCl47.6g,倒入刚才的那个烧杯中(PS:由于NaCl的量太多,一次称量不方便,所以分两次称量,且易于溶解)。往烧杯中加入Tris—HCl缓冲液100ml,最后加(吐温20)5ml,转入1000ml容量瓶中,在定容,转移即可。
TBST缓冲液的应用:
1.主要用于免疫组化和原位杂交,酶联免疫等实验中,清洗免疫印。
2.迹膜;
注意事项:
1.TBST缓冲液,PH7.2-7.5;
2.颜色为无色透明液体;
3.为了您的安全和健康,请穿实验服并戴防护手套操作;
1、 缓冲溶液对反应物的测定没有干扰
2、缓冲组分的浓度为1:1
3、有足够的缓冲容量
4、缓冲溶液的PH应在所需范围内
5、组成缓冲溶液的弱碱PKB和弱酸PKA应接近或等于所需的POH值或PH值(PH+POH=14)
配制
只要知道缓冲对的PH值,和要配制的缓冲液的pH值(及要求的缓冲液总浓度),就能按公式计算[盐]和[酸]的量。这个算法涉及对数换算,较麻烦,前人为减少后人的计算麻烦,已为我们总结出pH值与缓冲液对离子用量的关系并列出了表格。只要我们知道要配制的缓冲液的pH,经查表便可计算出所用缓冲剂的比例和用量。例如配制500nmpH5.8浓度为0.1M磷酸缓冲液。
经查表知pH5.8浓度为0.2M Na2HPO48.0毫升,而0.2M Na2HPO492.0毫升。依此可推论出配制100ml0.1M的磷酸缓冲液需要0.1M Na2HPO48.0毫升,而0.1M Na2HPO4需要92.0毫升。
计算好后,按计算结果准确称好固态化学成分,放于烧杯中,加少量蒸馏水溶解,转移入50ml容量瓶,加蒸馏水至刻度,摇匀,就能得到所需的缓冲液。
各种缓冲溶液的配制,均按表格按比例混合,某些试剂,必须标定配成准确浓度才能进行,如醋酸、氢氧化钠等。另外,所有缓冲溶剂的配制计量都能从以上的算式准确获得。
求采纳为满意回答。

