Bz-Ile-Glu-Gly-Arg-pNA.Acetate

• White to off-white lyophilized powder or crystalline solid.
• Hygroscopic; supplied in mannitol-bulked vials for stability.​

Source

• Chemically synthesized via solid-phase peptide synthesis with Fmoc protection, followed by benzoylation and pNA coupling.​
• Mixture of γ-OH (50%) and γ-methyl (50%) Glu forms for optimal solubility.

Molecular Weight and Structure

• C34H47N9O11 (acetate salt), 757.8 g/mol; base MW 741.3 (HCl form).
• N-benzoyl-Ile-Glu(γ-OR)-Gly-Arg-pNA; InChIKey ITXQNZUGLZNGBM-SFKTYRLKNA-N.​

Sugar Specificity

• Not applicable (peptide substrate); specific for trypsin-like endopeptidases cleaving after Arg.​

Biological Activity

• Hydrolyzed by Factor Xa (Km ~0.5-1 mM), releasing pNA (ε405 nm = 1.27 x 10^4 M⁻¹ cm⁻¹).
• Linear rate up to 4.8 µkat/L enzyme; optimal pH 7.5-8.5, 37°C.​

Purity and Microbial Contamination

• ≥98% HPLC purity; free pNA <0.05%; endotoxin-free for diagnostic use.
• Sterile, no microbial growth; heavy metals <10 ppm.​

Identity and Quality Control

• Verified by HPLC (single peak), MS (m/z 758 [M+H]+), amino acid analysis, and functional assay (ΔA405/min).
• Sequence confirmation via Edman degradation; CAS/IR spectra.​

Shelf Life and Storage

• 3-5 years at -20°C lyophilized; reconstitute in water/DMSO, use within 1 week at 4°C.
• Protect from moisture/light; stable in assay buffer for 8 hours.​

Application

• Factor Xa activity assays in plasma; anticoagulant potency (LMWH, DOACs); thrombin/thrombin time alternatives.
• Trypsin quantification; horseshoe crab coag enzyme; Factor VIII cofactor studies.​

Key Characteristics

• High specificity for Factor Xa over thrombin (100:1 ratio); sensitive detection limit 0.1 pM.​
• No interference from fibrinogen; automation-compatible; cost-effective for HTS.