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Equine D-Dimer,D2D ELISA Kit
Overview
Catalog No.: BZEK1758
Size: 96T
Range: 300 pg/ml - 4800 pg/ml
Sensitivity: 10 pg/ml
Storage and Expiration: Store at 2-8℃ for 6 months.
Application: For quantitative detection of D2D in equine serum, plasma, tissue homogenate or cell culture supernatant.
Introduction
D-dimer is a specific product of the degradation of fibrin clots that results from the action of 3 enzymes: (a) thrombin, generated from the activation of the coagulation cascade that converts fibrinogen into fibrin clots; (b) activated factor XIII that cross-links fibrin clots by means of covalent bonds between fibrin monomers; and (c) plasmin, the ultimate enzyme of fibrinolysis that degrades cross-linked fibrin (1,–,3). Monoclonal antibodies (MoAbs)2 raised against specific epitopes on D-dimer react with cross-linked fibrin, but not with fibrinogen degradation products or non–cross-linked fibrin degradation products, thus ensuring high specificity of the D-dimer as a biomarker of fibrin formation and stabilization (4). Many types of D-dimer assays have been developed that can be broadly divided into 3 categories.
Principle of the Assay
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- D2D antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- D2D antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the D2D amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of D2D can be calculated.
Kit components
1. One 96-well plate pre-coated with anti- equine D2D antibody
2. Standard: 0.5ml (9600 pg/ml)
3. Standard diluent buffer: 1.5 ml
4. Wash buffer (30×): 20 ml. Dilution: 1:30
5. Sample diluent buffer: 6 ml
6. HRP conjugated anti- equine D2D antibody (RTU): 6ml
7. Stop solution: 6 ml
8. TMB substrate A: 6ml
9. TMB substrate B: 6ml
10. Plate sealer: 2
11. Hermetic bag: 1
Material Required But Not Provided
1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Absorbent filter papers
8. Plastic or glass container with volume of above 1L
Other Information
Catalog No.: BZEK1758
Size: 96T
Range: 300 pg/ml - 4800 pg/ml
Sensitivity: 10 pg/ml
Storage and Expiration: Store at 2-8℃ for 6 months.
Application: For quantitative detection of D2D in equine serum, plasma, tissue homogenate or cell culture supernatant.
Introduction
D-dimer is a specific product of the degradation of fibrin clots that results from the action of 3 enzymes: (a) thrombin, generated from the activation of the coagulation cascade that converts fibrinogen into fibrin clots; (b) activated factor XIII that cross-links fibrin clots by means of covalent bonds between fibrin monomers; and (c) plasmin, the ultimate enzyme of fibrinolysis that degrades cross-linked fibrin (1,–,3). Monoclonal antibodies (MoAbs)2 raised against specific epitopes on D-dimer react with cross-linked fibrin, but not with fibrinogen degradation products or non–cross-linked fibrin degradation products, thus ensuring high specificity of the D-dimer as a biomarker of fibrin formation and stabilization (4). Many types of D-dimer assays have been developed that can be broadly divided into 3 categories.
Principle of the Assay
This kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. The purified anti- D2D antibody was pre-coated onto 96-well plates. And the HRP conjugated anti- D2D antibody was used as detection antibodies. The standards, test samples and HRP conjugated detection antibody were added to the wells subsequently, mixed and incubated, then, unbound conjugates were washed away with wash buffer. TMB substrates (A & B) were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the D2D amount of sample captured in plate. Read the O.D. absorbance at 450nm in a microplate reader, and then the concentration of D2D can be calculated.
Kit components
1. One 96-well plate pre-coated with anti- equine D2D antibody
2. Standard: 0.5ml (9600 pg/ml)
3. Standard diluent buffer: 1.5 ml
4. Wash buffer (30×): 20 ml. Dilution: 1:30
5. Sample diluent buffer: 6 ml
6. HRP conjugated anti- equine D2D antibody (RTU): 6ml
7. Stop solution: 6 ml
8. TMB substrate A: 6ml
9. TMB substrate B: 6ml
10. Plate sealer: 2
11. Hermetic bag: 1
Material Required But Not Provided
1. 37℃ incubator
2. Microplate reader (wavelength: 450nm)
3. Precise pipette and disposable pipette tips
4. Automated plate washer
5. ELISA shaker
6. 1.5ml of Eppendorf tubes
7. Absorbent filter papers
8. Plastic or glass container with volume of above 1L
Other Information
Please check datasheet for protocol.
