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This assay recognizes recombinant and natural mouse TpP

Mouse thrombus precursor protein (TpP) ELISA Kit


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Background

Thrombus precursor protein (TpP), which measures soluble fibrin, is a marker of active thrombosis. After thrombin cleaves fibrinopeptide A and B from fibrinogen, desAABB fibrin monomers are formed. These monomers polymerize into soluble TpP fibrin polymers, which serve as the immediate precursors to insoluble fibrin.
There was a significant association between greater levels of TpP and risk for the composite outcome of death, MI, recurrent ischemia leading to rehospitalization, or recurrent ischemia leading to urgent revascularization. Patients with ACS who present early after symptom onset have been shown to have significantly higher levels of TpP as compared to controls. Several smaller studies have evaluated TpP as a diagnostic tool and found that TpP appears to offer useful information for patients with ACS, particularly those patients who presented early after symptom onset. This marker of activated coagulation could potentially provide an additional pathophysiological axis for evaluation of the post-ACS patient.

Feature and Advantages

<PRINCIPLE OF THE ASSAY>
The microtiter plate provided in this kit has been pre-coated with an antibody specific to TpP. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for TpP and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain TpP, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of TpP in the samples is then determined by comparing the O.D. of the samples to the standard curve.

< SPECIFICITY>
This assay recognizes recombinant and natural mouse TpP. No significant cross-reactivity or interference was observed.

<DETECTION RANGE>
78 ng/ml-5000 ng/ml.
The standard curve concentrations used for the ELISA’s were 5000 ng/ml, 2500 ng/ml, 1250 ng/ml, 625 ng/ml, 312 ng/ml, 156 ng/ml, 78 ng/ml.

<SENSITIVITY>
The minimum detectable dose of mouse TpP is typically less than 19.5 ng/ml.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero.

Kit component

  Reagent Quantity
1. Assay plate 1
2. Standard 2
3. Sample Diluent 1 x 20 ml
4. Biotin-antibody Diluent 1 x 10 ml
5. HRP-avidin Diluent 1 x 10 ml
6. Biotin-antibodyt 1 x 120μl
7. HRP-avidin 1 x 120μl
8. Wash Buffer 1 x 20 ml (25×concentrate)
9. TMB Substrate 1 x 10 ml
10. Stop Solution 1 x 10 ml
Product List

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Species : 78 ng/ml-5000 ng/ml.
Assay range : 100 µL (Cell Culture Supernates, Plasma, Serum)
Assay sample volume : 6-8 hr.{3 hr


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