High Sensitive DBH ELISA Assay Kit For Canine Dopamine - Beta - Hydroxylase
Brand Name:BT Lab
Certification:CE, ISO9001:2005, MSDS
Model Number:Cat.No E0286Ca
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 Canine Dopamine-Beta-hydroxylase ELISA Assay Kit High
Sensitive DBH ELISA Kit
Cat.No E0286Ca
Referances
"Canine tyrosine hydroxylase (TH) gene and dopamine
beta-hydroxylase (DBH) gene: their sequences, genetic
polymorphisms, and diversities among five different dog breeds."
Takeuchi Y., Hashizume C., Chon E.M.H., Momozawa Y., Masuda K.,
Kikusui T., Mori Y.
J. Vet. Med. Sci. 67:861-867(2005)
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%
Standard Curve Range: 0.05ng/ml - 10ng/ml
Sensitivity: 0.025ng/ml
Size: 96 wells
Intended Use
This sandwich kit is for the accurate quantitative detection of
Canine Dopamine-Beta-hydroxylase (also known as DBH) in serum,
plasma, cell culture supernates, cell lysates, tissue homogenates.
Precautions
- Prior to use, the ELISA Assay 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 ELISA Assay Kit should not be used beyond the expiration date.
Reagent Preparation
All reagents should be brought to room temperature before use
Standard Reconstitute the 120μl of the standard (12.8ng/ml) with 120μl of
standard diluent to generate a 6.4ng/ml standard stock solution.
Allow the standard to sit for 15 mins with gentle agitation prior
to making dilutions. Prepare duplicate standard points by serially
diluting the standard stock solution (6.4ng/ml) 1:2 with standard
diluent to produce 3.2ng/ml, 1.6ng/ml, 0.8ng/ml and 0.4ng/ml
solutions. Standard diluent serves as the zero standard(0 ng/ml).
Any remaining solution should be frozen at -20°C and used within
one month. Dilution of standard solutions suggested are as follows:
| 6.4ng/ml | Standard No.5 | 120μl Original Standard + 120μl Standard Diluent |
| 3.2ng/ml | Standard No.4 | 120μl Standard No.5 + 120μl Standard Diluent |
| 1.6ng/ml | Standard No.3 | 120μl Standard No.4 + 120μl Standard Diluent |
| 0.8ng/ml | Standard No.2 | 120μl Standard No.3 + 120μl Standard Diluent |
| 0.4ng/ml | Standard No.1 | 120μl Standard No.2 + 120μl Standard Diluent |
| Standard Concentration | Standard No.5 | Standard No.4 | Standard No.3 | Standard No.2 | Standard No.1 |
| 12.8ng/ml | 6.4ng/ml | 3.2ng/ml | 1.6ng/ml | 0.8ng/ml | 0.4ng/ml |
Wash Buffer Dilute 20ml of Wash Buffer Concentrate 25x into deionized or
distilled water to yield 500 ml of 1x Wash Buffer. If crystals have
formed in the concentrate, mix gently until the crystals have
completely dissolved.
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-DBH
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.
High Sensitive DBH ELISA Assay Kit For Canine Dopamine - Beta - Hydroxylase
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