本公司生产的 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。
| 来源（Source） | 大肠杆菌重组表达 | | 分子量（Molecular Weight） | 26kDa | | 外观（Appearance） | 无菌液体 | | 保存液（Storage Buffer） | 50mM This-HCl , 100mM NaCl, 0.1mM MnCl₂, 0.1mM EGTA, 2mM dithiothreitol (DTT), 0.01%Brij35, 50%(v/v) glycerol, pH7.5 | | 酶浓度（Enzyme Concentration） | 70U/μL | | 纯度（Purity） | ≥95% | | 活性定义（Activity Definition） | 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 |
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组分和说明
| L747583-CG20KU | Component | 20KU | 5*20KU | Storage | | L747583A | Lambda Protein Phosphatase (70U/µl) | 300μl | 5*300μl | -20℃. Avoid freeze/thaw cycle. | | L747583B | 10× Lambda Protein Phosphatase Buffer | 1ml | 5*1ml | -20℃. Avoid freeze/thaw cycle. | | L747583C | 10× MnCl₂ (10mM MnCl₂) | 1ml | 5*1ml | -20℃. Avoid freeze/thaw cycle. |
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产品应用
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
产品优势
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。
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。
保存条件：
-20ºC 保存，1 年有效，≤0℃运输。
注意事项：
（1）反复冻融会导致 Lambda Protein Phosphatase 酶活性降低。如果希望在-80℃冻存，建议适当分装后保存，以方便后续使用。在-20℃存放时不会结冻，但在-80℃冻存时会结冻。
（2）10mM 钒离子(vanadate ions)能抑制 Lambda Protein Phosphatase 活性约 90%，50mM EDTA 可以抑制 Lambda Protein Phosphatase 活性约 95%，1% Triton X-100、0.4% Nonidet P-40、0.025% Tween 20、0.5M NaCl、0.1mM ATP、10μg/ml pepstatin A、10μg/ml leupeptin, 10μ
g/ml aprotinin, 0.5mM PMSF、1mM benzamidine 不影响 Lambda Protein Phosphatase 的酶活性。
（3）需要注意的是不同蛋白脱磷酸化反应的速率是不同的，该酶的最佳反应温度是 30℃。某些样品的粗提液用该酶进行去磷酸化处理时，请注意用尽可能短的时间进行处理，同时添加适量蛋白酶抑制剂以减少目的蛋白的降解。
（4）本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
（5）为了您的安全和健康，请穿实验服并戴一次性手套操作。
Lambda Protein Phosphatase (λ Protein Phosphatase) manufactured by our company is a Mn²⁺-dependent recombinant purified protein phosphatase. It catalyzes dephosphorylation of phosphorylated serine, threonine, tyrosine and histidine residues. This product is a 6×His-tagged recombinant λ phosphatase recombinantly expressed in Escherichia coli. The enzyme is encoded by open reading frame ORF221 of bacteriophage λ, and the encoded protein consists of 221 amino acid residues. Lambda Protein Phosphatase is applied for protein dephosphorylation, enabling research on the correlation between protein phosphorylation and protein activity/structure, as well as validation of the specificity of phospho-site antibodies.
| Source | Recombinantly expressed in E. coli
| | Molecular Weight | 26kDa | | Appearance | Sterile liquid
| | Storage Buffer | 50mM This-HCl , 100mM NaCl, 0.1mM MnCl₂, 0.1mM EGTA, 2mM Dithiothreitol (DTT), 0.01% Brij35, 50%(v/v) Glycerol, pH7.5 | | Enzyme Concentration | 70U/μL | | Purity | ≥95% | | Activity Definition | One unit (U) is defined as the quantity of enzyme that hydrolyzes 1 nmol of p-nitrophenyl phosphate within 1 minute at 30 °C and pH 7.5. |
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Components & Specifications
| L747583 | Component | 20KU | 5*20KU | Storage | | L747583A | Lambda Protein Phosphatase (70U/µl) | 300μl | 5*300μl | -20℃. Avoid freeze/thaw cycle. | | L747583B | 10× Lambda Protein Phosphatase Buffer | 1ml | 5*1ml | -20℃. Avoid freeze/thaw cycle. | | L747583C | 10× MnCl₂ (10mM MnCl₂) | 1ml | 5*1ml | -20℃. Avoid freeze/thaw cycle. |
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Product Applications:
Catalyze dephosphorylation of phosphorylated serine, threonine, tyrosine and histidine residues on target proteins.
Product Advantages:
Lambda Protein Phosphatase mediates protein dephosphorylation, which facilitates research on the correlation between protein phosphorylation and protein activity/structure. It can also be applied to verify the specificity of phospho-site-specific antibodies.
Instructions for Use:
1. Lambda Protein Phosphatase exhibits variable catalytic efficiency against different substrate proteins. Optimization of enzyme dosage and incubation time is recommended for individual experiments. The reaction system must contain 1× Lambda Protein Phosphatase Buffer supplemented with 1× MnCl₂ (final concentration: 1 mM).
2. Standard dephosphorylation reference system: In a total reaction volume of 50 μL, 100 U of Lambda Protein Phosphatase can remove approximately 100% phosphate groups from 5 μM monophosphorylated protein within 30 minutes.
Precautions:
1. Repeated freeze-thaw cycles will reduce the activity of Lambda Protein Phosphatase. If long-term storage at −80°C is required, aliquot the enzyme into smaller portions in advance for convenient subsequent use. The reagent remains liquid at −20 °C but will freeze at −80°C.
2. 10 mM vanadate ions inhibit approximately 90% of Lambda Protein Phosphatase activity, and 50 mM EDTA inhibits roughly 95% enzyme activity.
The following reagents do not interfere with enzyme activity: 1% Triton X-100, 0.4% Nonidet P-40, 0.025% Tween 20, 0.5 M NaCl, 0.1 mM ATP, 10 μg/mL pepstatin A, 10 μg/mL leupeptin, 10 μg/mL aprotinin, 0.5 mM PMSF, 1 mM benzamidine.
3. The dephosphorylation rate varies greatly among different substrate proteins. The optimal reaction temperature for this enzyme is 30°C. When treating crude lysate samples with this enzyme, minimize incubation time and add an appropriate cocktail of protease inhibitors to reduce degradation of target proteins.
4. This product is for scientific research use by trained professionals only. It must not be used for clinical diagnosis or therapy, food or pharmaceutical manufacturing, nor stored in residential premises.
5. Wear a lab coat and disposable nitrile/latex gloves throughout all operations for personal safety and health.
ZJ26F0838717
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