Pay in installments of $58.75 with
,
and
Shipping Estimate
USA
- USA
- CAN
- USA
- CAN
Ships within 48 hours · Estimated delivery Jul 25 - Jul 30
For Your Every Summer RSVP, with Code: SUMMER15
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. |
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) |
Shipping Notes
- Free Standard Shipping on $100+ Orders to the USA.
- Except Preorder products are shipped in 48 hours.
- Delivery to the USA:
- Standard Shipping : 3-10 business days
- If time is of the essence, please consider selecting expedited delivery for faster service.
Exchange/Return Notes
- We offer a 30-day return/exchange service after receiving.
- Final sale items are not eligible for returns or exchanges.
- To process your return/exchange, please contact us at [email protected]
- Please click here for more details>>> Return & Exchange Policy