Pefachrome® tPA, Chromogenic Substrate for tPA

Application:
Highly sensitive chromogenic peptide substrate for tissue-type plasminogen activator (tPA). Determination of tPA activity for research, in-process and quality control. Pefachrome. tPA shows different sensitivity to native single-chain tPA (sc-tPA) and its fully active two-chain form (tc-tPA). This allows a differential determination of each molecular form. 

Formula: CH₃SO₂-D-CHA-Gly-Arg-pNA · AcOH 

Principle:
CH₃SO₂-D-CHA-Gly-Arg-pNA + E => CH₃SO₂-D-CHA-Gly-Arg-OH + pNA + E
E = Enzyme (tc-tPA or sc-tPA) 

MW: 658.9 

Kinetic data:

	KM	Vmax
sc-tPA	0.286 mM	6.95 nmol/µg tPA/min
tc-tPA	0.167 mM	33.9 nmol/µg tPA/min

 

Storage:
May be used by the expiry date given on the label when stored unopened, protected from moisture, in the dark, 2-8°C. Avoid contamination of the reagents by micro-organisms. Shipment of product does not require cooling during the time of transportation.

Material required but not provided:
Buffer, reference material

Buffer: 50 mM Tris-Imidazole buffer pH 8.4, 300 mM NaCl, 37 °C

Assay:
Suggested protocol for the determination of native single-chain tPA (sc-tPA):
1.780 ml buffer
0.020 ml Enzyme (approx. 75’000 IU /ml)
0.200 ml Pefachrome^®tPA (2 mM in water)
=> Determination of ∆OD/min at 405 nm

Suggested protocol for the determination of two-chain tPA (tc-tPA):
1.750 ml buffer
0.050 ml Enzyme (approx. 120’000 IU /ml)
0.200 ml Pefachrome^®tPA (4 mM in water)
=> Determination of ∆OD/min at 405 nm

Category: Research use only

Type: Substrate

Product Availability: Germany, Israel and Egypt

Manufacturer: DSM Nutritional Products Ltd. Branch Pentapharm, Switzerland