Human D-Dimer,D2D ELISA Kit

Code CSB-E05175h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Human D-Dimer (D2D) ELISA Kit is suitable for qualitatively determining D2D concentrations in multiple biological fluids, including human serum and plasma in vitro. D-dimer is the final degradation product of cross-linked fibrin generated during plasma coagulation. Yumeng Yao etc. demonstrated that D-dimer expression is commonly elevated in patients infected with SARS-CoV-2. D-dimer levels correlate with disease severity and are a reliable prognostic marker for in-hospital mortality in patients admitted for COVID-19.

This kit uses the quantitative sandwich-based enzyme immunoassay technique to measure the amount of human D2D in the sample. Standards and samples are respectively added to the microplate wells pre-coated with anti-human D2D antibody. Biotin-labeled D2D antibody, HRP-avidin, and TMB substrate are pipped into the microplate in turn. D2D in the sample or standard is captured by the immobilized antibody and then binds to the biotinylated D2D monoclonal antibody. And the biotin on the biotinylated D2D antibody binds to the avidin on the enzyme label, forming immune complexes. The color renders blue after the addition of TMB substrate. The addition of the stop solution into the wells immediately turns the blue solution into a yellow one. The concentration of D2D in the samples is directly proportional to the OD value (450nm). Each manufactured lot of this ELISA kit was quality tested for criteria such as sensitivity, specificity, precision, linearity, recovery, and lot-to-lot consistency. See the manual for more information on validation.

Target Name D-Dimer,D2D
Abbreviation D2D
Species Homo sapiens (Human)
Sample Types serum, plasma
Detection Range 15.6 ng/mL-1000 ng/mL
Sensitivity 3.9 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cardiovascular
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human D2D in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 87  
Range % 80-95  
1:2 Average % 96  
Range % 91-100  
1:4 Average % 102  
Range % 92-110  
1:8 Average % 92  
Range % 84-98  
Recovery
The recovery of human D2D spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 89 80-96  
EDTA plasma (n=4) 95 90-105  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
1000 2.695 2.498 2.597 2.457  
500 2.289 2.138 2.214 2.074  
250 1.544 1.486 1.515 1.375  
125 0.813 0.783 0.798 0.658  
62.5 0.415 0.401 0.408 0.268  
31.2 0.279 0.274 0.277 0.137  
15.6 0.217 0.211 0.214 0.074  
0 0.141 0.138 0.140    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human D2D antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard --- Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled D2D antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody solution.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) --- Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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