SKU: 227154152

PLK1 GST&His Tag Protein, Human

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Description

PLK1 GST&His Tag Protein, HumanProduct 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

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)



 

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SKU: 227154152

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LanaB
Carnegie, US
★★★★★ 5
Ends in a cliffhanger
Format: Kindle
Thankfully, the next book is already available. Roxy Collins is the queen of omegaverse slow burn. This particular series is omegaverse with wolf shifters. Thoroughly enjoyed this book! Lots of suspense and trying to figure out what is true and what's really going on. I loved it! Pretty spicy once the action starts. Heats are pretty much always spicy. There is a little bit of male interaction, but not full-on shmex. Definitely recommend.
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Reviewed in the United States on April 29, 2025
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Dolores Evans
Waukegan, US
★★★★★ 5
Epic!!!!
Format: Kindle
Absolutely addicted from the first page. The twists and turns, world building and character development all come together for the perfect Omegaverse. And let's not forget that spice because knots are life and these are spectacular. Running for book #2 because that cliffhanger was torturous.
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Reviewed in the United States on August 17, 2024
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Ruth Ann Burt
Whiting, US
★★★★★ 5
Great book
Format: Kindle
I absolutely feel in love with all 4 characters!!! The bedroom scenes were 🌋🌡🔥🔥🔥. I couldn't put this book down!!! I'm hooked for the whole series Book 2 here I come!!!!! Its a fun easy book and story to read!!
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Reviewed in the United States on October 4, 2024
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Danyelle
Lexington, US
★★★★★ 4
Fun with a late blooming omega
Format: Kindle
I like this book. The story is fun, cute, and sexy. There's just a little drama, some excellent, steamy scenes, and a fairly good relationship building storyline. I especially like how all the main characters are a bit older than the usual 20 somethings I tend to see in this kind of book. Having said that, I wish there were more descriptions of the places, as well as the food in the fancy restaurant. I enjoyed the cocktails at the club, so I missed that kind of detail when Gray took Madison on a dinner date. I also wish there had been more interaction between Lucas and Madison, and Lucas and Rian. It felt a bit lopsided, with a focus on Rian, Madison, and Gray. I wish it had been proofread - there are a lot of typos, but nothing too distracting.
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Reviewed in the United States on September 12, 2022
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Jennifer G
Carnegie, US
★★★★★ 3
Madison Deserved Better
Format: Kindle
Madison was a beta...except she wasn't any longer. She was a late presenting Omega. And she was struggling. She was tall and thin, not tiny and curvy. She was opinionated. She was everything an Omega was not. After suffering through her first heat, her friends took her to Ardor, a club where Omegas came to safely find Alphas. She's not expecting much but then she connects with a sexy beta. And when she meets his Alphas, they set her body on fire. Maybe, she's found her no-strings-attached heat pack. Maybe, she's found something more. I could not connect with the characters in this book, so their story never resonated with me. And there was no love story; there was sex. Grey made it clear from the beginning that he had a true love and it was his beta boy, Rian. He went so far as to reassure Rian “Say the word, I’ll never touch her again. Lucas can put the babies in her. I only need you, beta boy”. So, Madison was there for babies, no emotions needed. Nice. No, thank you. I want the Omega to be the center of their world, not an incubator. Lucas and Rian weren't any better. After her heat, they let her leave. Not one of them made her feel valued. No one gave her a reason to stay or even offered a cuddle. And the sex didn't even come across as mind-blowing. Madison deserved better.
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Reviewed in the United States on March 11, 2025

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