T3 Sandwich Bovine ELISA Kits Customized Tri - iodothyronine Immunoassay Test Kit 96 Wells
Brand Name:BT Lab
Certification:CE, ISO9001:2015, MSDS
Model Number:Cat.No E0215Bo
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
Bovine T3 Sandwich ELISA Kit Customized Tri-iodothyronine
Immunoassay Test Kit 96 Wells Plate
Cat.No E0215Bo
Standard Curve Range: 0.03ng/ml - 9ng/ml
Sensitivity: 0.01ng/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.
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%
Intended Use
This sandwich kit is for the accurate quantitative detection of
Bovine Tri-iodothyronine (also known as T3) in serum, plasma, cell
culture supernates, cell lysates, tissue homogenates.
Assay Principle
This ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The
plate has been pre-coated with Bovine T3 antibody. T3 present in
the sample is added and binds to antibodies coated on the wells.
And then biotinylated Bovine T3 Antibody is added and binds to T3
in the sample. Then Streptavidin-HRP is added and binds to the
Biotinylated T3antibody. 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 Bovine T3.
The reaction is terminated by addition of acidic stop solution and
absorbance is measured at 450 nm.
Reagent Provided
| Components | Quantity |
| Standard Solution (9.6ng/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 (30x) | 20ml x1 |
| Biotinylated Bovine T3 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
Precautions
- Prior to use, the kit and sample should be warmed naturally to room temperature 30 minutes.
- This instruction must be strictly followed in the experiment.
- Once the desired number of strips has been removed, immediately reseal the bag to protect the remain from deterioration. Cover all reagents when not in use.
- Make sure pipetting order and rate of addition from well-to-well when pipetting reagents.
- Pipette tips and plate sealer in hand should be clean and disposable to avoid cross-contamination.
- Avoid using the reagents from different batches together.
- Substrate solution B is sensitive to light, don’t expose substrate solution B to light for a long time.
- Stop solution contains acid. Please wear eye, hand and skin protection when using this material. Avoid contact of skin or mucous membranes with kit reagent.
- The kit should not be used beyond the expiration date.
Assay Procedure
1. Prepare all reagents, standard solutions and samples as
instructed. Bring all reagents to room temperature before use. The
assay is performed at room temperature.
2. Determine the number of strips required for the assay. Insert
the strips in the frames for use. The unused strips should be
stored at 2-8°C.
3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution
contains biotinylated antibody.
4. Add 40μl sample to sample wells and then add 10μl anti-T3
antibody to sample wells, then add 50μl streptavidin-HRP to sample
wells and standard wells (Not blank control well). Mix well. Cover
the plate with a sealer. Incubate 60 minutes at 37°C.
5. Remove the sealer and wash the plate 5 times with wash buffer.
Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1
minute for each wash. For automated washing, aspirate all wells and
wash 5 times with wash buffer, overfilling wells with wash buffer.
Blot the plate onto paper towels or other absorbent material.
6. Add 50μl substrate solution A to each well and then add 50μl
substrate solution B to each well. Incubate plate covered with a
new sealer for 10 minutes at 37°C in the dark.
7. Add 50μl Stop Solution to each well, the blue color will change
into yellow immediately.
8. Determine the optical density (OD value) of each well
immediately using a microplate reader set to 450 nm within 10
minuets after adding the stop solution.
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.
T3 Sandwich Bovine ELISA Kits Customized Tri - iodothyronine Immunoassay Test Kit 96 Wells
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