| 货号 (SKU) | 包装规格 | 是否现货 | 价格 | 数量 |
|---|---|---|---|---|
| L747583-20KU |
20KU |
期货  |
| |
| L747583-100KU |
100KU |
期货 |
|
| 产品名称 | Lambda Protein Phosphatase |
|---|---|
| 别名 | λ蛋白磷酸酶 |
| 规格或纯度 | ≥95%, Lambda Protein Phosphatase存储液组成为：50mM This-HCl (pH7.5)，100mM NaCl，0.1mM MnCl2，0.1mM EGTA，2mM dithiothreitol (DTT)，0.01%Brij35，50%(v/v)甘油。10× Lambda Protein Phosphatase Buffer组成为：500mM Hepes (pH7.5)，100mM NaCl，20mM DTT，0.1% Brij35。10× MnCl2组成为：10mM M |
| 产品介绍 |
阿拉丁生产的Lambda Protein Phosphatase (重组λ蛋白磷酸酶)，即λ Protein Phosphatase，是一种锰离子(Mn2+)依赖的重组表达纯化的磷酸蛋白酶，可以对磷酸化的丝氨酸、苏氨酸、酪氨酸和组氨酸残基进行脱磷酸反应。本产品是大肠杆菌重组表达的带6×His标签的重组λ磷酸酶。该磷酸酶由λ噬菌体开放阅读框ORF221所编码，编码产物共包含221个氨基酸残基。Lambda Protein Phosphatase可以用于蛋白的去磷酸化，从而可以用于研究蛋白磷酸化与其活性和结构的关系，以及验证蛋白磷酸化位点抗体的特异性等。酶活性单位定义：在30℃ pH7.5条件下反应1分钟，能够水解1nmol p-nitrophenyl phosphate所需的酶量定义为一个活性单位。阿拉丁生产的Lambda Protein Phosphatase酶活性鉴定结果参考图1。图1. Lambda Protein Phosphatase酶活测定实验图。左图：pNP标准曲线。右图：Lambda Protein Phosphatase按照30、40、50、80、100、200、500倍稀释后，在50µl反应体系中的酶量分别为0.27µg、0.20µg、0.16µg、0.10µg、0.08µg、0.04µg、0.016µg，并与100mM pNPP在1×Lambda Protein Phosphatase Buffer和1×MnCl2中在30℃反应5min后，加入12.5µl 3M NaOH终止反应，测定产生的pNP在405nm吸光度，并根据pNP标准曲线，计算Lambda Protein Phosphatase酶活性。不同批次检测出来的酶活性会略有差别。酶分子量大小：约26kDa。纯度：纯度大于95%。Lambda Protein Phosphatase存储液组成为：50mM This-HCl (pH7.5)，100mM NaCl，0.1mM MnCl2，0.1mM EGTA，2mM dithiothreitol (DTT)，0.01%Brij35，50%(v/v)甘油。10× Lambda Protein Phosphatase Buffer组成为：500mM Hepes (pH7.5)，100mM NaCl，20mM DTT，0.1% Brij35。10× MnCl2组成为：10mM MnCl2。失活：在50mM EDTA存在的条件下，65℃加热1小时可使Lambda Protein Phosphatase失活。 失活或抑制： 在50mM EDTA存在的条件下，65℃加热1小时可使Lambda Protein Phosphatase失活。使用说明： 1.Lambda Protein Phosphatase对不同蛋白的脱磷酸化反应有不同活性，所以在实际应用中，我们建议对酶浓度和反应时间进行优化。反应体系中，反应缓冲液为1× Lambda Protein Phosphatase Buffer并且同时加入1× MnCl2 (最终浓度为1mM)。2.以一般的去磷酸化反应为例：在50μl反应体系中，100U的Lambda Protein Phosphatase在30分钟内可以除去相当于5μM的磷酸化的蛋白(单磷酸化)中约100%的磷酸。Aladdin's Lambda Protein Phosphatase is a Mn2+-dependent protein phosphatase with activity towards phosphorylated serine, threonine, tyrosine and histidine residues. It can be used to dephosphorylate proteins to study the relationship between protein phosphorylation and its activity and structure, and to verify the specificity of antibodies to protein phosphorylation sitesThis product is a recombinant lambda phosphatase with 6×His tag expressed in E. coli. It is the 221 amino-acid product of the ORF221 open reading frame on bacteriaphage lambda.Definition of enzyme activity unit Precautions： Repeated freeze-thaws will reduce the enzyme activity of Lambda Protein Phosphatase. When stored at -80℃, it should be aliquoted.Vanadate ions at 10mM can inhibit Lambda Protein Phosphatase activity by about 90%, and 50mM EDTA can inhibit Lambda Protein Phosphatase activity by about 95%. The enzyme activity of Lambda Protein Phosphatase is not affected by 1% Triton X-100, 0.4% Nonidet P-40, 0.025% Tween 20, 0.5 M NaCl, 0.1mM ATP, 10μg/ml pepstatin A, 10μg/ml leupeptin, 10μg/ml aprotinin, 0.5mM PMSF, or 1mM benzamidine.Vanadate ions at 10mM can inhibit Lambda Protein Phosphatase activity by about 90%, and 50mM EDTA can inhibit Lambda Protein Phosphatase activity by about 95%. The enzyme activity of Lambda Protein Phosphatase is not affected by 1% Triton X-100, 0.4% Nonidet P-40, 0.025% Tween 20, 0.5 M NaCl, 0.1mM ATP, 10μg/ml pepstatin A, 10μg/ml leupeptin, 10μg/ml aprotinin, 0.5mM PMSF, or 1mM benzamidine.This product has an optimal reaction temperature of 30℃, and its dephosphorylation rate varies for different proteins. When treating the crude extracts of some samples with this enzyme, the duration time for the treatment should be as short as possible and appropriate amount of protease inhibitors should be also added to minimize the degradation of target proteins.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use： 1. The activity of Lambda Protein Phosphatase varies for different proteins. Empirically, optimal enzyme concentrations and incubation times should be determined for each particular protein substrate. Beside the 1X Lambda Protein Phosphatase Buffer, MnCl2 should be supplemented in the reaction mix to a final concentration of 1mM.2. Take a general dephosphorylation reaction as an example: in a 50μl reaction mix, 100U Lambda Protein Phosphatase can remove about 100% of phosphate groups from 5μM mono-phosphorylated proteins within 30 minutes. |
| 浓度 | Lambda Protein Phosphatase存储液组成为：50mM This-HCl (pH7.5)，100mM NaCl，0.1mM MnCl2，0.1mM EGTA，2mM dithiothreitol (DTT)，0.01%Brij35，50%(v/v)甘油。10× Lambda Protein Phosphatase Buffer组成为：500mM Hepes (pH7.5)，100mM NaCl，20mM DTT，0.1% Brij35。10× MnCl2组成为：10mM MnCl2。 |
|---|---|
| 储存温度 | -20°C储存 |
| 运输条件 | 超低温冰袋运输 |
| 稳定性与储存 | -20℃保存，一年有效。如果希望保存更长时间，可以适当分装后-80℃冻存。 |
| 分子类型 | 蛋白质 |
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