PLK1 GST&His Tag Protein, Human

Serine/threonine-protein kinase PLK1,Polo-like kinase 1,Serine/threonine-protein kinase 13 (STPK13)

Datasheet COA

价格￥2,800.00 （供应商现货： 3-5个工作日）

货号 UA085052

规格

10μg50μg100μg

数量

产品介绍

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产品规格

物种

Human
分子别名

Serine/threonine-protein kinase PLK1, Polo-like kinase 1, Serine/threonine-protein kinase 13 (STPK13)
Accession

P53350
表达序列

Met1-Ser603 with GST Tag at the N-Terminus and His Tag at the C-Terminus
表达宿主

Baculovirus-InsectCells
分子量

100-130kDa (Reducing)
纯度

＞90% by SDS-PAGE
活性

The specific activity of PLK1 GST&His Tag Protein, Human was equivalent to >20 nmol/min/mg as per activity assay protocol.
标记

Unconjugated
标签

GST Tag, His Tag
性状

Liquid
缓冲体系

50mM sodium phosphate, PH 7.0, 300mM NaCl, 0.25mM DTT, and 10% glycerol
储存条件

Stable for 12 months upon stored at -80℃ from the date of receipt. And avoid repeated freeze-thaws cycles.
文献引用

1. Structural regulation of PLK1 activity: implications for cell cycle function and drug discovery. (2025). Cancer Gene Therapy, 32, 608-621.
2. Targeting polo-like kinase 1: advancements and future directions in anti-cancer drug discovery. (2024). Expert Opinion on Drug Discovery, 19(10), 1153-1157.
3. PLK1 inhibition by Shikonin attenuates oral cancer growth through β-catenin-mediated regulation of EMT and apoptosis. (2026). Cellular Signalling, 138, 112262.

背景介绍

PLK1 (Polo-like Kinase 1) is a highly conserved serine/threonine protein kinase that serves as a master regulator of cell division. It belongs to the Polo-like kinase family, which comprises five members (PLK1-5), with PLK1 being the most extensively studied. The protein is encoded by the PLK1 gene located on chromosome 16p12.2 and consists of 603 amino acids with a molecular weight of approximately 66 kD .Structural Organization: PLK1 features a unique dual-domain architecture. The N-terminus contains a conserved catalytic kinase domain (KD) responsible for phosphorylating substrates. The C-terminus contains a unique polo-box domain (PBD) composed of two polo-box motifs (PB1 and PB2) that function as a phosphopeptide recognition module. The PBD binds to substrates that have been "primed" by phosphorylation at specific Ser-pThr-(Pro) motifs by other kinases (notably CDK1), thereby determining substrate specificity and subcellular localization

操作步骤

实验方法
实验原理：使用ADP-Glo™激酶检测试剂盒进行PLK1活性测定，该试剂盒用于定量检测PLK1反应产生的ADP量。具体步骤如下：首先加入ADP-Glo™试剂以终止激酶反应并消耗剩余ATP；随后加入激酶检测试剂，将ADP转化为ATP，并通过荧光素酶/荧光素反应体系检测新合成的ATP。
实验材料
1.Kinase assay buffer(5X): 200 mM Tris-HCl, pH 7.4, 100 mM MgCl2 and 0.5 mg/mL BSA, 250 μM DTT
2.Kinase assay buffer(1X): 40 mM Tris-HCl, pH 7.4, 20 mM MgCl2, 0.1 mg/mL BSA, 50 μM DTT
3.PLK1 GST Tag & His Tag Protein, Human
4.ADP-Glo Kinase Assay (UA, Catalog # UA070101)
5.底物: PLKtide peptide (Sinobiological, Catalog # P41-5)
6.Solid white multi-well plate (384-well plate) (Corning, Catalog #3572)
7.读板器（PerkinElmer）
实验步骤
1.配制底物/ATP混合液，操作如下（以25 μM为例）：
Sample Name
Amount (μL)
10 mM ATP Solution
1
Kinase Assay Buffer III (5x)
79
Substrate at 1 mg/mL
80

2.用激酶检测缓冲液（1x）将PLK1稀释至20 µg/mL, 10 µg/mL和5 µg/mL，并在 384 孔板的每个孔中加入3 µL。
3.通过向每个孔中加入2 µL微升步骤1中配制的检测系统来启动反应。设置一个仅加入 3 µL激酶检测缓冲液（1x）的检测系统作为空白对照。反应总体积为5 µL。将反应在室温（22–25℃）下孵育40分钟。
4.向反应完成的体系中加入5 µL ADP-Glo Reagent，短暂混匀后在室温（22–25℃）下孵育 40分钟。
5.加入10 µL Detection Reagent，将板在室温（22–25℃）下孵育30分钟。
6.分别使用终点模式读取化学发光信号。
7.计算比活性。
标准曲线
1.在激酶检测缓冲液（1×）中将ATP和ADP稀释至25 μM。
2.按下表所示，混合25 μM ATP和25 μM ADP，配制ATP+ADP混合液，并在384孔板的每个孔中加入5 μL。
Well Number
1
2
3
4
5
6
7
8
9
10
11
12
25μM ADP (μL)
100
80
60
40
20
10
5
4
3
2
1
0
25μM ATP (μL)
0
20
40
60
80
90
95
96
97
98
99
100

3.向反应完成的孔中加入5 μL ADP-Glo试剂，短暂混匀，在室温（22-25°C）下孵育40分钟。
4.加入10 μL检测试剂，在室温（22-25°C）下孵育30分钟。
5.在终点模式下分别读取化学发光信号。
6.检测光信号并建立转换曲线。
Specific Activity (pmol/min/μg) =
ATP (pmol)-Blank
Incubation time(min) ×amount of enzyme (μg)

生物活性
- The specific activity of PLK1 GST&His Tag Protein, Human was equivalent to >20 nmol/min/mg as per activity assay protocol.
电泳
- 2μg (R: reducing condition, N: non-reducing condition).