Fluorogenic Substrates

Transcription

1 PENTAPHARM Fluorogenic Substrates Pentapharm Ltd. offers a broad spectrum of fluorogenic peptide substrates for research, inprocess and quality control, employing the fluorogenic group AMC (7-Amino-4-methylcoumarin). Optical characteristics of AMC: Absorption maximum wavelength: λ Abs. : 342 nm Emission maximum wavelength: λ Em. : 440 nm Pefafluor FXa Highly sensitive fluorogenic peptide substrate for factor Xa. Formula: CH 3 SO 2 -D-CHA-Gly-Arg-AMC AcOH MW: k cat : s -1 K M : 0.22 mm Solubility: Up to 10 mm in dest. H 2 O Suggested protocol for the determination of factor Xa activity: ml 50 mm Tris-HCl ph 7.4, 100 mm NaCl, 0.5% HSA ml Factor Xa (KORDIA, 11.8 nm in NaCl) ml Pefafluor FXa (1.5 mm in water) Package size: Vial containing 10 µmol Code: Pentapharm Ltd, Engelgasse 109, P.O. Box, CH-4002 Basel / Switzerland 1/5

2 Pefafluor TH Sensitive fluorogenic peptide substrate for thrombin. Determination of thrombin activity for research, in-process and quality control. Formula: H-D-CHA-Ala-Arg-AMC 2AcOH MW: K M : 1.93 µm k cat : 53.9 s -1 Suggested protocol for the determination of thrombin in microplates: Stock solutions: Thrombin (3 NIH U/ml in 300 mm NaCl) ml bovine thrombin solution ml 0.05 M Tris/HCl, 300 mm NaCl, 0.5% HSA, ph ml Pefafluor TH (2 mm) => determination of fluorescence emission at 460 nm Excitation wavelength 360 nm / Emission wavelength 460 nm Package size: Vial containing 25 mg Code: Pefafluor tpa Highly sensitive fluorogenic peptide substrate for tissue-type plasminogen activator (tpa). Formula: CH 3 SO 2 -D-Phe-Gly-Arg-AMC AcOH MW: 673 k cat : 11.0 s -1 K M : 0.14 mm Suggested protocol for the determination of sc-tpa activity: ml 100 mm HEPES ph 8.0, 154 mm NaCl, 0.1% HSA ml sc-tpa (0.5 µm) ml Pefafluor tpa (1.0 mm in water) Package size: Vial containing 25 mg Code: Pentapharm Ltd, Engelgasse 109, P.O. Box, CH-4002 Basel / Switzerland 2/5

3 Pefafluor upa Highly sensitive fluorogenic peptide substrate for urokinase (upa). Formula: Bz-β-Ala-Gly-Arg-AMC AcOH MW: v max : µmol. l -1. s -1 K M : mm Suggested protocol for the determination of urokinase: ml 50 mm Tris-HCl ph 8.0, 100 mm NaCl ml upa (Ribosepharm, 850 IU/mg) ml Pefafluor upa (0.5 mm in water) Package size: Vial containing 10 µmol Code: Pefafluor PCa Sensitive fluorogenic peptide substrate for activated protein C. Formula: Pyr-Pro-Arg-AMC AcOH MW: k cat : 62.0 s -1 K M : 0.56 mm Suggested protocol for the determination of activated protein C: ml 50 mm Tris-HCl ph 8.0, 100 mm NaCl, 0.5% HSA ml activated protein C (KORDIA, 46 nm) ml Pefafluor PCa (10 mm in water) Package size: Vial containing 25 mg Code: Pentapharm Ltd, Engelgasse 109, P.O. Box, CH-4002 Basel / Switzerland 3/5

4 Pefafluor PCa3342 Sensitive chromogenic peptide substrate with significantly improved selectivity for activated protein C. Formula: Pyr-CHG-Arg-AMC AcOH MW: k cat : 17.0 s -1 K M : 1.60 mm Suggested protocol for the determination of activated protein C: ml 50 mm Tris-HCl ph 8.0, 100 mm NaCl, 0.5% HSA ml Activated protein C (KORDIA, 46 nm) ml Pefafluor PCa3342 (1.5 mm in water) Package size: Vial containing 25 mg Code: Pefafluor LAL Highly sensitive fluorogenic peptide substrate for the determination of bacterial endotoxins. Formula: CH 3 SO 2 -D-HHT-Gly-Arg-AMC AcOH MW: Package size: Vial containing 25 mg Code: Pentapharm Ltd, Engelgasse 109, P.O. Box, CH-4002 Basel / Switzerland 4/5

5 Pefafluor FIXa10148 Fluorogenic peptide substrate for factor IXa with improved sensitivity. Determination of factor IXa activity for in-process and quality control of factor IX preparations. formula: Mes-(D)-CHG-Gly-Arg-AMC. AcOH MW: V max : 28.1 µmol/min K M : 0.23 mm (determined in the presence of 33% ethylene glycol) Suggested protocol for an assay in microplates: Buffer: 50 mm Tris ph 7.4, 100 mm NaCl, 5 mm CaCl 2, 40% ethylene glycol ml Buffer ml Pefafluor FIXa10148 (10 mm in water) ml FIXa: human factor IXaβ, Enzyme Research, final conc µg/ml Note: The sensitivity of Pefafluor FIXa10148 is significantly increased in the presence of 33% ethylene glycol. Package size: Code: Pefafluor FIXa3688 Highly sensitive fluorogenic peptide substrate for factor IXa. Formula: H-(D)-Leu-PHG-Arg-AMC. 2AcOH MW: V max : 1.12 µmol/min K M : mm Suggested protocol for the determination of factor IXa activity: ml Pefafluor FIXa3668 (10 mm in water) ml Buffer (50 mm Tris-HCl ph 7.4, 100 mm NaCl, 5 mm CaCl 2, 0.5% HSA) ml FIXa: human factor IXaβ, Enzyme Research, final conc µg/ml Package size: Code: [Data sheet / 5802] [Fluorogenic Substrates.doc] Registered Trademark of Pentapharm Ltd. Pentapharm Ltd, Engelgasse 109, P.O. Box, CH-4002 Basel / Switzerland 5/5