0.11ng/Ml Sensitivity ELISA Assay Kit ADAMTS19 High Precision For Research
Brand Name:BT Lab
Certification:CE, ISO9001:2005, MSDS
Model Number:Cat.No E3489Hu
Minimum Order Quantity:Negotiation
Delivery Time:1-3 business days, bulk order within one week
Place of Origin:Shanghai, China
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Product Details
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Product Details
Customized Human A Disintegrin and Metalloproteinase with
Thrombospondin Motifs 19 ELISA Assay Kit
Cat.No E3489Hu
Standard Curve Range: 0.2ng/ml - 60ng/ml
Sensitivity: 0.11ng/ml
Size: 96 wells
Storage: Store the reagents at 2-8°C. For over 6-month storage refer to
the expiration date keep it at -20°C. Avoid repeated thaw cycles.
If individual reagents are opened it is recommended that the kit be
used within 1 month.
*This product is for research use only, not for use in diagnosis
procedures. It’s highly recommend to read this instruction entirely
before use.
Reagent Provided
| Components | Quantity |
| Standard Solution (64ng/ml) | 0.5ml x1 |
| Pre-coated ELISA Plate | 12 * 8 well strips x1 |
| Standard Diluent | 3ml x1 |
| Streptavidin-HRP | 6ml x1 |
| Stop Solution | 6ml x1 |
| Substrate Solution A | 6ml x1 |
| Substrate Solution B | 6ml x1 |
| Wash Buffer Concentrate (25x) | 20ml x1 |
| Biotinylated human ADAMTS19 Antibody | 1ml x1 |
| User Instruction | 1 |
| Plate Sealer | 2 pics |
| Zipper bag | 1 pic |
Material Required But Not Supplied
- 37°C±0.5°C incubator
- Absorbent paper
- Precision pipettes and disposable pipette tips
- Clean tubes
- Deionized or distilled water
- Microplate reader with 450 ± 10nm wavelength filter
Precision
Intra-Assay Precision (Precision within an assay) Three samples of
known concentration were tested on one plate to assess intra-assay
precision.
Inter-Assay Precision (Precision between assays) Three samples of
known concentration were tested in separate assays to assess
inter-assay precision.
CV(%) = SD/mean x 100
Intra-Assay: CV<8%
Inter-Assay: CV<10%
Assay Principle
This elisa assay kit is an Enzyme-Linked Immunosorbent Assay
(ELISA). The plate has been pre-coated with human ADAMTS19
antibody. ADAMTS19 present in the sample is added and binds to
antibodies coated on the wells. And then biotinylated human
ADAMTS19 Antibody is added and binds to ADAMTS19 in the sample.
Then Streptavidin-HRP is added and binds to the Biotinylated
ADAMTS19 antibody. After incubation unbound Streptavidin-HRP is
washed away during a washing step. Substrate solution is then added
and color develops in proportion to the amount of human ADAMTS19.
The reaction is terminated by addition of acidic stop solution and
absorbance is measured at 450 nm.
Specimen Collection
Serum Allow serum to clot for 10-20 minutes at room temperature.
Centrifuge at 2000-3000 RPM for 20 minutes.
Plasma Collect plasma using EDTA or heparin as an anticoagulant.
Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C
within 30 minutes of collection.
Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for
approximately 20 minutes. When collecting pleuroperitoneal fluid
and cerebrospinal fluid, please follow the procedures
above-mentioned.
Cell Culture Supernatant Collect by sterile tubes when examining secrete components.
Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect
the supernatants carefully. When examining the components within
the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the
cell concentration of approximately 1 million/ml. Damage cells
through repeated freeze-thaw cycles to let out the inside
components. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
Tissue and other body fluids Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly
and weigh before homogenization. Mince tissues and homogenize them
in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or
freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
Note
- Sample concentrations should be predicted before being used in the assay. If the sample concentration is not within the range of the standard curve, users must contact us to determine the optimal sample for their particular experiments.
- Samples to be used within 5 days should be stored at 2-8°C. Samples should be aliquoted or must be stored at -20°C within 1 month or -80°C within 6 months. Avoid repeated freeze thaw cycles.
- Samples should be brought to room temperature before starting the assay.
- Centrifuge to collect sample before use.
- Samples containing NaN3 can’t be tested as it inhibits the activity of Horse Radish Peroxidase (HRP).
- Collect the supernatants carefully. When sediments occurred during storage, centrifugation should be performed again.
- Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.
*Sample can't be diluted with this kit. Owing to the the material we
use to prepare the kit, the sample matrix interference may falsely
depress the specificity and accuracy of the assay.
Summary
1. Prepare all reagents, samples and standards.
2. Add sample and ELISA reagent into each well. Incubate for 1 hour
at 37°C.
3. Wash the plate 5 times.
4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.
5. Add stop solution and color develops.
6. Read the OD value within 10 minutes.
Calculation of Result
Construct a standard curve by plotting the average OD for each
standard on the vertical (Y) axis against the concentration on the
horizontal (X) axis and draw a best fit curve through the points on
the graph. These calculations can be best performed with
computer-based curve-fitting software and the best fit line can be
determined by regression analysis.
0.11ng/Ml Sensitivity ELISA Assay Kit ADAMTS19 High Precision For Research
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