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D-dimer is a protein formed by two D fragments of the fibrin protein joined by a cross-link. D-dimer is one of several fibrin degradation product (FDP) formed by the degradation of a blood clot by fibrinolysis. It is used in the diagnosis of the blood disorder disseminated intravascular coagulation and in the diagnosis of thrombosis.
This kit is designed for the quantitative measurement of Fibrinogen D-Dimer protein in Mouse Plasma.
Please note that this kit is also available as a CLIA Kit abx490569
Research Articles on Mouse D-Dimer ELISA Kit
||Optimal dilutions/concentrations should be determined by the end user.
||Shipped at 4 °C. Upon receipt, store the kit according to the storage instruction in the kit's manual.
||The validity for this kit is 6 months.
||The stability of the kit is determined by the rate of activity loss. The loss rate is less than 5% within the expiration date under appropriate storage conditions. To minimize performance fluctuations, operation procedures and lab conditions should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same user throughout.
||0.617 ng/ml - 50 ng/ml
||< 0.284 ng/ml
||This kit is based on competitive enzyme-linked immuno-sorbent assay technology. An antibody is pre-coated onto a 96-well plate. Standards, test samples, and biotin-conjugated reagent are added to the wells and incubated. A competitive inhibition reaction takes place between the biotin-labelled D-Dimer and the unlabelled- D-Dimer on the pre-coated antibody. The HRP-conjugated reagent is then added, and the whole plate is incubated. Unbound conjugates are removed using wash buffer at each stage. TMB substrate is used to quantify the HRP enzymatic reaction. After TMB substrate is added, only wells that contain sufficient D-Dimer will produce a blue coloured product, which then changes to yellow after adding the acidic stop solution. The intensity of the color yellow is inversely proportional to the D-Dimer amount bound on the plate. The OD is measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of D-Dimer can be calculated.
- Pre-coated 96-Well Microplate
- Standard Diluent Buffer
- Wash Buffer
- Detection Reagent A
- Detection Reagent B
- Diluent A
- Diluent B
- TMB Substrate
- Stop Solution
- Plate Sealer
|Material Required But Not Provided
- 37°C incubator
- Multi and single channel pipettes and sterile pipette tips
- Squirt bottle or automated microplate washer
- 1.5 ml tubes
- Distilled water
- Absorbent filter papers
- 100 ml and 1 liter graduated cylinders
- Microplate reader (wavelength: 450 nm)
- ELISA Shaker
- Serum: Samples should be collected into a serum separator tube. Coagulate the serum by leaving the tube undisturbed in a vertical position overnight at 4°C or at room temperature for up to 60 minutes. Centrifuge at approximately 1000 × g for 20 min. Analyze the serum immediately or aliquot and store at -20°C or -80°C.
- Plasma: Collect plasma using heparin or EDTA as an anticoagulant. Centrifuge for 15 minutes at 1000 × g within 30 minutes of collection. Assay immediately or aliquot and store at -20°C or -80°C. Avoid hemolysis and high cholesterol samples.
- Tissue homogenates: The preparation of tissue homogenates will vary depending upon tissue type - this is just an example. Rinse tissues with ice-cold PBS to remove the excess of blood. Weigh before homogenization. Finely mince tissues and homogenize with a tissue homogenizer on ice in PBS and sonicate the cell suspension. Centrifuge the homogenates at 5000 × g for 5 min and collect the supernatant. Assay immediately or aliquot and store at -20°C.
- 1) Standard: Prepare the standard with the recommended volume of Standard Diluent Buffer, to make the standard solution. Then use the Standard Diluent buffer to carry out serial dilutions of the standard solution, as instructed in the Protocol.
- 2) Wash Buffer: Dilute the concentrated Wash Buffer with distilled water, as instructed in the Protocol.
- 3) Detection Reagent Preparation: Calculate the total volume of working solution required. Dilute Detection Reagent A and Detection Reagent B with Diluent A and Diluent B, respectively, at 1:100.
- 1) Set standard, test samples and control wells.
- 2) Aliquot 50 μl of diluted standard into the standard wells.
- 3) Aliquot 50 μl of Standard Diluent buffer into the control (zero) well.
- 4) Aliquot 50 μl of diluted samples into the sample wells.
- 5) Immediately aliquot 50 μl of Detection Reagent A to each well. Incubate for 1 hr at 37 °C.
- 7) Aliquot 100 μl of Detection Reagent B to each well. Incubate for 30 mins at 37 °C.
- 9) Aliquot 90 μl of TMB Substrate to each well. Incubate for 10-20 mins at 37 °C.
- 10) Aliquot 50 μl of Stop Solution.
- 11) Measure the OD at 450 nm.
- Equilibrate the kit components and samples to room temperature (18 - 25 °C) before use. It is recommended to plot a standard curve for each test.
- 1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample at least in duplicate.
- 2. Add 50 µL of each standard, control and sample into the appropriate wells. 3. Remove the cover and discard the liquid.
- 4. Immediately aliquot 50 μl of Detection Reagent A working solution. Seal the plate with a cover and incubate for 1 h at 37°C.
- 5. Remove the cover and discard the solution. Wash the plate 3 times with 1X Wash Buffer.
- 6. Add 100 µL of Detection Reagent B working solution into each well, seal and incubate at 37°C for 30 min.
- 7. Discard the solution and wash the plate 5 times with wash buffer as explained in previous step.
- 8. Aliquot 90 μl of TMB Substrate into each well. Seal the plate with a cover and incubate at 37°C for 10-20 min. Avoid exposure to light. The incubation time is for reference use only, the optimal time should be determined by end user. Do not exceed 30 min.
- 9. Add 50 µL of Stop Solution to each well. Read at 450 nm immediately.
||This assay is competitive, therefore there is an inverse correlation between D-Dimer concentration in the sample and the absorbance measured. Create a graph with the log of the standard concentration (y-axis) and average absorbance measured (x-axis). Apply a best fit trendline through the standard points. The D-Dimer concentration of the samples can be interpolated from the standard curve.
||Intra-assay Precision (Precision within an assay): 3 samples with low, medium and high levels of C-X-C Motif Chemokine 9 / MIG (CXCL9) were were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, medium and high levels of C-X-C Motif Chemokine 9 / MIG (CXCL9) were tested on 3 different plates, 8 replicates in each plate.
CV (%) = (Standard Deviation / mean) × 100
||Shipped within 5-7 working days.
||This product is for research use only. The range and sensitivity is subject to change. Please contact us for the latest product information. For accurate results, sample concentrations must be diluted to mid-range of the kit. If you require a specific range, please contact us in advance or write your request in your order comments. Please note that our ELISA and CLIA kits are optimised for detection of native samples, rather than recombinant proteins. We are unable to guarantee detection of recombinant proteins, as they may have different sequences or tertiary structures to the native protein.