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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
2-8℃
- 保质期:
6个月
- 英文名:
Soil Polyphenoloxidase(S-PPO) Activity Assay Kit
- 库存:
99
- 供应商:
北京索莱宝科技有限公司
- CAS号:
BC0110-50T/24S
- 规格:
50T/24S
土壤多酚氧化酶(S-PPO)活性检测试剂盒说明书 详见附件
可见分光光度法
货号: BC0110
规格: 50T/48S
产品组成:使用前请认真核对试剂体积与瓶内体积是否一致,有疑问请及时联系索莱宝工作人员。
| 试剂名称 | 规格 | 保存条件 |
| 试剂一 | 粉剂×2瓶 | 2-8℃保存 |
| 试剂二 | 液体10 mL×1瓶 | 2-8℃保存 |
| 试剂三 | 液体 100 mL×1瓶(自备) | 2-8℃保存 |
| 标准液 | 液体10 mL×1瓶 | 2-8℃保存 |
产品说明:
S-PPO主要来源于土壤微生物、植物根系分泌物及动植物残体分解释放,催化土壤中芳香族化合物氧化成醌,醌与土壤中蛋白质、氨基酸、糖类、矿物等物质反应生成有机质和色素,完成土壤芳香族化合物循环,用于土壤环境修复。
S-PPO能够催化邻苯三酚产生紫色没食子素,后者在430nm有特征光吸收。
注意:实验之前建议选择2-3个预期差异大的样本做预实验。如果样本吸光值不在测量范围内建议稀释或者增加样本量进行检测。
- 样本处理(可适当调整待测样本量,具体比例可以参考文献)
测定步骤
1、可见分光光度计预热30min以上,调节波长至430nm,试剂三调零。
2、标准液稀释:用0.5mol/L HCl溶液将标准液稀释至0.2、0.1、0.05、0.025、0.0125、0.00625、0.003125、0 mg/mL。试验中需标准品1000µL。
3、标准液稀释表:
| 序号 | 稀释前浓度(mg/mL) | 标准液体积(µL) | 0.5mol/L HCl体积(µL) | 稀释后浓度(mg/mL) |
| 1 | 0.2 | 1000 | 0 | 0.2 |
| 2 | 0.2 | 1000 | 1000 | 0.1 |
| 3 | 0.1 | 1000 | 1000 | 0.05 |
| 4 | 0.05 | 1000 | 1000 | 0.025 |
| 5 | 0.025 | 1000 | 1000 | 0.0125 |
| 6 | 0.0125 | 1000 | 1000 | 0.00625 |
| 7 | 0.00625 | 1000 | 1000 | 0.003125 |
| 8 | - | 0 | 1000 | 0 |
4、标准曲线的建立:以0 mg/mL标准液调零,取1mL稀释好的标准液于1mL玻璃比色皿中在430nm处测定吸光值A,根据吸光度(x)和浓度(y,mg/mL)做出标准曲线。
5、操作表:
| 试剂名称 | 测定管 |
| 风干土样(g) | 0.05 |
| 试剂一(μL) | 500 |
| 振荡混匀,30℃水浴反应1h | |
| 试剂二(μL) | 200 |
| 试剂三(μL) | 1750 |
S-PPO活性计算
根据标准曲线,将样本吸光值A(x)带入公式中,计算出样本浓度y(mg/mL)。
单位的定义:每天每g土样中产生1mg紫色没食子素定义为一个酶活力单位。
S-PPO活性(U/g 土样)=y×V提取相÷W÷T=840×y
T:反应时间,1h=1/24d;V提取相:1.75mL;W:样本质量,0.05g。
相关发表文献:
- B Li,Y Ding, X Tang,et al. MTA1 promotes the invasion and migration of pancreatic cancer cells potentially through the HIF-α/VEGF pathway. Journal of Receptor and Signal Transduction Research. August 2018;(IF2.998)
[1] Montgomery M W, Sgarbieri V C. Isoenzymes of banana polyphenol oxidase[J]. Phytochemistry, 1975, 14(5-6): 1245-1249.
[2] Dogan S, Dogan M. Determination of kinetic properties of polyphenol oxidase from Thymus (Thymus longicaulis subsp. chaubardii var. chaubardii)[J]. Food chemistry, 2004, 88(1): 69-77.

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