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PLK1 GST&His Tag Protein, Human

PLK1 GST&His Tag Protein, Human

Catalog Number: UA085052 Brand: UA BIOSCIENCE
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Product Details

Product Specification


Species Human
Synonyms Serine/threonine-protein kinase PLK1, Polo-like kinase 1, Serine/threonine-protein kinase 13 (STPK13)
Accession P53350
Amino Acid Sequence

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

Expression System Baculovirus-InsectCells
Molecular Weight

100-130kDa (Reducing)

Purity >90% by SDS-PAGE
Conjugation Unconjugated
Tag His Tag, GST Tag
Physical Appearance Liquid
Storage Buffer

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

Stability & Storage

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

Reference

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.

Background

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

Protocol

Assay protocol

Principle: The PLK1 assay is performed using the ADP-GloTM Kinase Assay kit which quantifies the amount of ADP produced by the PLK1 reaction. The ADP-GloTM Reagent is added to terminate the kinase reaction and to deplete the remaining ATP, and then the Kinase Detection Reagent is added to convert ADP to ATP and to measure the newly synthesized ATP using luciferase/luciferin reaction.

Materials

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.Substrate: PLKtide peptide (Sinobiological, Catalog # P41-58)

6.Solid white multi-well plate (384-well plate) (Corning, Catalog #3572)

7.Plate Reader (PerkinElmer)

Produce

1.Prepare a substrate/ATP mixture as follows (25 μM example).

Sample Name

Amount (μL)

10 mM ATP Solution

1

Kinase Assay Buffer III (5x)

79

Substrate at 1 mg/mL

80


 

2. Dilute the PLK1 to 20 µg/mL, 10 µg/mL and 5 µg/mL in Kinase Assay Buffer (1x) and dispense 3 μL into each well of a 384-well plate.

3. Initiate the reaction by adding 2 μL of the detection system prepared in Step 1 to each well. Include a detection system with 3 μL Kinase Assay Buffer (1x) as Blank. The reaction volume is 5 μL.

4. Incubate the reaction at room temperature (22–25℃) for 40 minutes.

5.Add 5 μL of ADP-Glo Reagent to the completed reaction, mix briefly and incubate for 40 minutes at room temperature (22–25 ℃).

6.Add 10 μL of Detection Reagent and incubate the plate for 30 minutes at room temperature (22–25 ℃).

7.Read at luminescence, respectively in endpoint mode.

8.Calculate specific activity.

• Standard Curve

1.Dilute the ATP and ADP to 25 μM in Kinase Assay Buffer (1x).

2.Mix 25 μΜ ATP and 25 μM ADP to form an ATP+ADP solution provided below and dispense 5 μL into each well of a 384-well plate.

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.Add 5 μL of ADP-Glo Reagent to the completed reaction, mix briefly and incubate for 40 minutes at room temperature (22–25 ℃).

4.Add 10 μL of Detection Reagent and incubate the plate for 30 minutes at room temperature (22–25 ℃).

5.Read at luminescence, respectively in endpoint mode.

6.Detect optical signals and establish conversion curves.

Specific Activity (pmol/min/μg) =

ATP (pmol)-Blank

Incubation time(min) ×amount of enzyme (μg)



 

Picture

Bioactivity

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

SDS-PAGE

2μg (R: reducing condition, N: non-reducing condition).