Microplate-based antithrombin activity bioassay for Shuxuetong Injection through aptamer-thrombin capturing coupled with chromogenic substrate hydrolysis

2021 ◽  
pp. 106099
Author(s):  
Xiaofang Wang ◽  
Dan Yan
Keyword(s):  
Chromogenic Substrate ◽  
Antithrombin Activity ◽  
Substrate Hydrolysis

The Presence of Antithrombin Activity in Platelets

1979 ◽  
Author(s):  
E. E. Czapek ◽  
H. C. Kwaan
Keyword(s):  
Antithrombin Iii ◽  
Human Platelet ◽  
Platelet Rich Plasma ◽  
Clot Formation ◽  
Chromogenic Substrate ◽  
Antithrombotic Activity ◽  
Platelet Poor Plasma ◽  
Antithrombin Activity ◽  
At Iii ◽  
Platelet Pellet

The platelet has been implicated as the nidus around which clot formation occurs. We therefore decided to investigate whether any antithrombotic activity was associated with platelets. Thrombin activity was measured using the chromogenic substrate S-2238. Freshly prepared human platelet rich plasma (PRP) and platelet poor plasma (PPP) were found to have similar amounts of antithrombin activity. This antithrombin activity could be easily removed from a platelet pellet by repeated washing. However, sonicating PRP resulted in the appearance of additional antithrombin activity. After 6 minutes of sonication, the antithrombin activity of PRP decreased to 35% of presonication levels, whereas the antithrombin activity of PPP remained 100%. When subjected to Laurell Immunoelectrophoresis using an antibody to antithrombin III (AT-III), a greater amount of antigen was detected in PRP than in PPP. We conclude that platelets contain antithrombin material which is probably AT-III.

Automated Enzyme Assay of Antithrombin

1980 ◽  
Vol 17 (4) ◽  
pp. 183-184 ◽  
Author(s):  
S A Cederholm-Williams
Keyword(s):  
Enzyme Assay ◽  
Chromogenic Substrate ◽  
Antithrombin Activity ◽  
Automated Assay ◽  
Relevant Range

The conditions for the automated assay of plasma antithrombin activity over the clinically relevant range of 25–250% using the chromogenic substrate S2238 and the Gilford 3800 enzyme rate analyser are reported.

Measurement of antithrombin activity by thrombin-based and by factor Xa-based chromogenic substrate assays

2000 ◽  
Vol 11 (2) ◽  
pp. 127-135 ◽  
Author(s):  
H. Beeck ◽  
D. Nagel ◽  
G. Pindur ◽  
I. Scharrer ◽  
A. Preiss ◽  
...  
Keyword(s):  
Factor Xa ◽  
Chromogenic Substrate ◽  
Antithrombin Activity

Measurement of antithrombin activity by thrombin-based and by factor Xa-based chromogenic substrate assays

2000 ◽  
Vol 11 (2) ◽  
pp. 127-135 ◽  
Author(s):  
H. Beeck ◽  
D. Nagel ◽  
G. Pindur ◽  
I. Scharrer ◽  
A. Preiss ◽  
...  
Keyword(s):  
Factor Xa ◽  
Chromogenic Substrate ◽  
Antithrombin Activity

The Presence of Antithrombin Activity in Platelets

1979 ◽  
Author(s):  
E.E. Czapek ◽  
H.C. Kwaan
Keyword(s):  
Antithrombin Iii ◽  
Human Platelet ◽  
Platelet Rich Plasma ◽  
Clot Formation ◽  
Chromogenic Substrate ◽  
Antithrombotic Activity ◽  
Platelet Poor Plasma ◽  
Antithrombin Activity ◽  
At Iii ◽  
Platelet Pellet

The platelet has been implicated as the nidus around which clot formation occurs. We therefore decided to investigate whether any antithrombotic activity was associated with platelets. Thrombin activity was measured using the chromogenic substrate S-2238. Kreshly prepared human platelet rich plasma (PRP) and platelet poor plasma (PPP) were found to have similar amounts of antithrombin activity. This antithrombin activity could be easily removed from a platelet pellet by repeated washing. However, sonicating PRP resulted in the appearance of additional antithrombin activity. After 6 minutes of sonlcation, the antithrombin activity of PRP decreased to 35% of presonication levels, whereas the antithrombin activity of PPP remained 100%. When subjected to Laurell immunoelectrophoresis using an antibody to antithrombin III(AT-III), a greater amount of antigen was detected in PRP than in PPP. We conclude that platelets contain antithrombin material which is probably AT-III.

Factor XII Determinations in the Presence and Absence of Phospholipid Antibodies

1998 ◽  
Vol 79 (01) ◽  
pp. 87-90 ◽  
Author(s):  
D. W. Jones ◽  
M. Winter ◽  
M. J. Gallimore
Keyword(s):  
Lower Limit ◽  
Lupus Anticoagulant ◽  
Factor Xii ◽  
Chromogenic Substrate ◽  
Plasma Samples ◽  
Presence And Absence ◽  
Lower Limit Of Normal

SummaryFactor XII (FXII) levels were determined in plasma samples from 29 normal donors, 10 patients with inherited FXII deficiency (all lupus anticoagulant [LA] negative) and 67 LA positive patients, using clotting (FXIIct), chromogenic substrate (FXIIcs) and immunochemical (FXIIag) assays. Excellent correlations were obtained in the three FXII assays with the LA negative samples and between the FXIIcs and FXIIag assays in the LA positive samples. Correlations between both the FXIIcs and FXIIag with FXIIct in the LA positive patients were poor. Of 67 LA positive samples studied, 25 (37.3%) showed lower values in the FXIIct assay; 13 (19.4%) of these patients were pseudo FXII deficient with values of FXII below the lower limit of normal.These results indicate that a diagnosis of FXII deficiency can be made inappropriately in the presence of phospholipid antibodies and that such a diagnosis should not be made by FXIIct assay alone.

Visualization of von Willebrand Factor Multimers by Enzyme-Conjugated Secondary Antibodies

1986 ◽  
Vol 55 (02) ◽  
pp. 276-278 ◽  
Author(s):  
F Brosstad ◽  
Inge Kjønniksen ◽  
B Rønning ◽  
H Stormorken
Keyword(s):  
Von Willebrand Factor ◽  
Chromogenic Substrate ◽  
Agarose Electrophoresis ◽  
Secondary Antibodies ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Beta Galactosidase

SummaryA method for visualization of the multimeric forms of von Willebrand Factor (vWF) in plasma and platelets is described. The method is based upon: 1) Separation of the vWF multimers by SDS-agarose electrophoresis, 2) Subsequent blotting of the vWF multimers onto nitrocellulose, 3) Immunolocalization and visualization of the vWF pattern by the sequential incubation of the blot with a) primary vWF antiserum, b) peroxidase- or beta-galactosidase-conjugated secondary antibodies and a relevant chromogenic substrate.

Plasma Thrombin Assay using a Chromogenic Substrate in Disseminated Intravascular Coagulation due to Snake Bite Envenomation

1979 ◽  
Vol 41 (03) ◽  
pp. 544-552 ◽  
Author(s):  
R P Herrmann ◽  
P E Bailey
Keyword(s):  
Disseminated Intravascular Coagulation ◽  
Degradation Products ◽  
Snake Bite ◽  
Chromogenic Substrate ◽  
Coagulation Parameters ◽  
Fibrinogen Degradation Products ◽  
Intravascular Coagulation

SummaryUsing the chromogenic substrate, Tos-Gly-Pro-Arg-pNA-HCL (Chromozym TH, Boehringer Mannheim) plasma thrombin was estimated in six cases of envenomation by Australian elapid snakes. All patients manifested findings chracteristic of defibrination due to envenomation by these snakes. Fibrin-fibrinogen degradation products were grossly elevated, as was plasma thrombin in all cases.Following treatment with antivenene, all abnormal coagulation parameters returned rapidly towards normal by 24 hours and plasma thrombin disappeared.

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