THE MICROFOCUS ASSAY SYSTEM (MFAS) - A SIMPLE QUANTITATIVE ASSAY TO IDENTIFY INHIBITORY DRUGS TARGETED AGAINST SPECIFIC ONCOGENES

1994 ◽  
Author(s):  
DB REARDON ◽  
YM XIE ◽  
MC HUNG
Keyword(s):  
Assay System ◽  
Quantitative Assay

Discovery of novel poly(ADP-ribose) glycohydrolase inhibitors by a quantitative assay system using dot-blot with anti-poly(ADP-ribose)

2010 ◽  
Vol 392 (4) ◽  
pp. 485-489 ◽  
Author(s):  
Naoyuki Okita ◽  
Daisuke Ashizawa ◽  
Ryo Ohta ◽  
Hideaki Abe ◽  
Sei-ichi Tanuma
Keyword(s):  
Assay System ◽  
Quantitative Assay ◽  
Dot Blot

Quantitative assay system for specific enzyme activity using antibody: the case of protease, subtilisin BPN′

1998 ◽  
Vol 66 (2-3) ◽  
pp. 157-163 ◽  
Author(s):  
Yoshiaki Miyota ◽  
Shingo Komada ◽  
Haruo Momose ◽  
Seiichi Taguchi
Keyword(s):  
Enzyme Activity ◽  
Assay System ◽  
Quantitative Assay ◽  
Specific Enzyme ◽  
Specific Enzyme Activity

The Significance of TFPI in Clotting Assays – Gomparison and Combination with other Anticoagulants

1993 ◽  
Vol 70 (03) ◽  
pp. 448-453 ◽  
Author(s):  
Ole Nordfang ◽  
Hanne I Kristensen ◽  
Sanne Valentin ◽  
Per Østergaard ◽  
Johnny Wadt
Keyword(s):  
Tissue Factor ◽  
Tissue Factor Pathway Inhibitor ◽  
Antithrombin Iii ◽  
Anticoagulant Activity ◽  
Assay System ◽  
Thrombin Inhibitor ◽  
Clotting Time ◽  
Normal Plasma ◽  
Tissue Factor Pathway ◽  
Plasma Heparin

SummaryThe anticoagulant activities of Tissue Factor Pathway Inhibitor (TFPI), heparin and hirudin were compared in intrinsic (APTT) and extrinsic (PT) activated clotting assays. In contrast to the thrombin inhibitor hirudin, heparin was 10 fold more potent in the APTT assay than in the PT assay, indicating that inhibition of intrinsic activation is important for the anticoagulant activity of heparin as measured in an APTT assay. TFPI was most potent in the PT assay and the effect of TFPI was most pronounced in the presence of other anticoagulants (heparin and hirudin). The activities of the two natural anticoagulants antithrombin III (ATIII) and TFPI were compared in a PT assay with very dilute tissue factor. In this assay system TFPI in normal plasma affected the clotting time more than ATIII in the plasma. However, when heparin was added ATIII was the major anticoagulant, but profound Prolongation of the clotting time was only seen when TFPI was also added. In an ATIII deficient plasma heparin did not augment the effect of TFPI, showing that the increased effect of TFPI in the presence of heparin is dependent on the anticoagulant activity of ATIII/heparin. The effect of TFPI at prolonged clotting times was also illustrated by the significant effect of blocking TFPI in the plasma from warfarin-treated patients. Thus TFPI is a major anticoagulant in normal plasma and the effect of TFPI is especially seen at prolonged clotting times.

A Comparative Ex Vivo Study of Plasminogen Activators and Proteases for Fibrinolytic Activity and Side Effects in Rabbits

1977 ◽  
Vol 37 (01) ◽  
pp. 154-161 ◽  
Author(s):  
B. A Janik ◽  
S. E Papaioannou
Keyword(s):  
Side Effects ◽  
Effective Dose ◽  
Fibrinolytic Activity ◽  
Ex Vivo ◽  
Plasminogen Activators ◽  
Assay System ◽  
Coagulation Parameters ◽  
Ex Vivo Assay

SummaryUrokinase, streptokinase, Brinase, trypsin, and SN 687, a bacterial exoprotease, have been evaluated in an ex vivo assay system. These enzymes were injected into rabbits and the fibrinolytic activity as well as other coagulation parameters were measured by in vitro techniques. Dose-response correlations have been made using the euglobulin lysis time as a measure of fibrinolytic activity and the 50% effective dose has been determined for each enzyme. Loading doses, equal to four times the 50% effective dose, were administered to monitor potential toxicity revealing that Brinase, trypsin, and SN 687 were very toxic at this concentration.Having established the 50% effective dose for each enzyme, further testing was conducted where relevant fibrinolytic and coagulation parameters were measured for up to two days following a 50% effective dose bolus injection of each enzyme. Our results have demonstrated that urokinase and streptokinase are plasminogen activators specifically activating the rabbit fibrinolytic system while Brinase, trypsin and SN 687 increase the general proteolytic activity in vivo.The advantages of this ex vivo assay system for evaluating relative fibrinolytic potencies and side effects for plasminogen activators and fibrinolytic proteases have been discussed.

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