Chinese Name: Thrombin Chromogenic Substrate S-2160

English Name: Bz-Phe-Val-Arg-pNA

Abbreviation: S-2160

CAS Number: 54799-93-8

Molecular Formula: C33H40N8O6

Molecular Weight: 644.73

Specifications: 5 mg / 100 mg / 250 mg (Custom packaging supported)

Form: Off-white crystalline powder

Product Number: G04040016

Stock: Made to order

Signal: 405 nm colorimetric (pNA, yellow)

Chinese Name: Thrombin Chromogenic Substrate S-2160

English Name: Bz-Phe-Val-Arg-pNA ∙ HCl

Abbreviation: S-2160

CAS Number: 38789-84-3

Molecular Formula: C33H41ClN8O6

Molecular Weight: 681.19

Specifications: 5 mg / 100 mg / 250 mg (Custom packaging supported)

Form: Off-white crystalline powder

Product Number: G04040017

Stock: Made to order

Signal: 405 nm colorimetric (pNA, yellow)

The S-2160 series substrates are built around the sequence Bz-Phe-Val-Arg as the enzymatic recognition core: thrombin performs specific cleavage at the arginine residue at the P1 position, and the C-terminal signal group is released, converting the catalytic event into an optical signal. The type of signal group determines the detection method:

• pNA (p-nitroaniline): turns yellow upon release, quantified by colorimetry at 405 nm;

• AMC (7-amino-4-methylcoumarin): emits fluorescence upon release, Ex 360-380 nm / Em 440-460 nm;

• 4MβNA (4-methoxy-β-naphthylamide): emits fluorescence upon release; 4MβNA has fluorescence characteristics distinct from AMC, suitable for specific detection systems and platform requirements;

The peptide sequence anchors substrate specificity, while the signal group defines the detection dimension – the same enzymatic core, three observational perspectives.

The S-2160 series is built around the Bz-Phe-Val-Arg substrate sequence as the core scaffold, with differences originating from two dimensions: signal group and salt form:

Table 1 GeneSeqTools S-2160 Series Substrates

Selection logic:

1. Colorimetric vs. Fluorescent – determined by detection sensitivity requirements

Table 2 Comparison of S-2160 Colorimetric and Fluorescent Substrate Characteristics

• Colorimetric substrates (pNA series):

Suitable for scenarios where equipment versatility and cost sensitivity are prioritized, the first choice for most routine assays.

• Fluorescent substrates (AMC / 4MβNA series):

Suitable for scenarios with low enzyme activity, requiring a wider dynamic range of signals or higher signal-to-noise ratio, such as thrombin activity detection, drug screening gradient experiments, etc.

2. Free base or hydrochloride – determined by operational mode

• Free base:

Complete literature data, suitable for routine laboratory solution preparation, ideal for early method development.

• HCl salt:

Superior dissolution rate and saturation concentration compared to the free base, suitable for the following scenarios:

◦ Automated solution preparation platforms requiring rapid dissolution and avoiding precipitation

◦ GMP production with higher requirements for preparation efficiency and batch consistency

◦ Need to prepare high-concentration stock solutions to accommodate high-throughput screening formats

3. Complementary relationship between 4MβNA and AMC

The spectral characteristics of 4MβNA differ from those of AMC, making it a complementary choice to AMC under equipment configuration limitations or specific methodological requirements.

1. Coagulation diagnostic reagent development

Used for antithrombin III detection systems, thrombin activity detection methods, and related IVD product development.

As a thrombin chromogenic substrate, S-2160 can be used to construct:

• Antithrombin III assay kits

• Thrombin activity assay kits

• Supporting reagents for automated coagulation analyzers

Helping coagulation diagnostic reagent developers solve core raw material needs in reagent formulation development, detection method establishment, and product performance validation.

2. Anticoagulant drug evaluation

Used for evaluating changes in thrombin activity and pharmacodynamic studies during the development of anticoagulant drugs such as heparins and novel oral anticoagulants.

By detecting the change in thrombin activity after drug action, it enables:

• Heparin activity evaluation

• High-throughput screening of anticoagulants

• Pharmacodynamic studies

S-2160 can be used for thrombin activity detection, with fluorescent forms (AMC/4MβNA) suitable for low-signal sample analysis and high-throughput screening platforms.

3. Serine protease research

Used for thrombin and related serine protease activity detection and associated enzymology studies.

By detecting the signal changes of released pNA/AMC/4MβNA from S-2160 under protease action, it supports:

• Determination of thrombin kinetic parameters (Km, Vmax, kcat)

• Substrate specificity studies of serine proteases

• Exploration of coagulation pathway mechanisms

Helping research institutions solve core tool requirements for enzymatic characterization of thrombin and related serine proteases and substrate specificity studies.

✅ Classic thrombin substrate with mature application foundation

• Classic sequence: Designed based on the Bz-Phe-Val-Arg recognition sequence, long-term application in thrombin activity detection.

• Rich literature accumulation: Over 50 years of application research foundation, widely used in coagulation research and method development.

• Reduced development risk: Mature application systems help shorten method establishment and validation cycles.

✅ Clear substrate recognition characteristics, supporting detection system establishment

• Specificity is clear: Designed for thrombin detection, suitable for quantitative analysis of thrombin activity.

• Method development support: Clear substrate reaction characteristics, useful for establishing and optimizing thrombin activity detection systems.

• Good data reproducibility: Conducive to establishing stable, repeatable detection workflows.

✅ Adapted to coagulation and anticoagulation research systems

• Wide application coverage: Supports antithrombin III functional testing, heparin activity evaluation, and anticoagulant drug development.

• From research to IVD raw material: Meets multi-level needs from basic research, drug development to diagnostic reagent development.

• Supports platform expansion: Same recognition sequence covers both colorimetric and fluorescence platform applications.

✅ Domestic R&D, independent and controllable supply chain

• Cost advantage: For the same quality, cost is significantly lower than imported brands, helping customers reduce costs and improve efficiency.

• Controllable delivery time: Independently produced by GeneSeqTools, stable delivery reduces import procurement cycles and supply risks.

• Multiple specifications available: Available in 5 mg / 100 mg / 250 mg, with custom packaging supported, meeting needs from research trial to industrial mass production.