DA ELISA Kit,Absin_specification/price/image

Product Detail
Company Profile

Product Specification

Usage

Equipment required for the experiment:
1. Microplate reader (450nm)
2. High-precision pipette and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL
3. 37℃ constant temperature box
4. Distilled water or deionized water

Sample processing and requirements:
Serum: Place the whole blood sample collected in the serum separation tube at room temperature for 2 hours or at 4℃ overnight, then centrifuge at 1000×g for 20 minutes and take the supernatant. Alternatively, store the supernatant at -20℃ or -80℃, but avoid repeated freezing and thawing. Plasma: Collect the specimen using EDTA or heparin as an anticoagulant. Centrifuge at 1000×g for 15 minutes at 2-8°C within 30 minutes of collection. Remove the supernatant for testing. Alternatively, store the supernatant at -20°C or -80°C, but avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge at 1000×g for 20 minutes. Remove the supernatant for testing. Pre-Test Preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and allow it to equilibrate to room temperature. 2. Preparation of Gradient Standard Working Solution: Add 1 mL of Universal Diluent to the lyophilized standard. Let stand for 15 minutes to completely dissolve, then gently mix (concentration 1000 pg/mL). Then, dilute to the following concentrations: 1000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.25 pg/mL, 15.625 pg/mL, and 0 pg/mL. Serial Dilution Method: Add 500 μL of Universal Diluent to each of seven EP tubes. Pipette 500 μL of the 1000 pg/mL standard working solution into the first EP tube and mix thoroughly to make a 500 pg/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details.

3. Preparation of Biotin-Antibody Working Solution: Centrifuge the concentrated Biotin-Antibody at 1000×g for 1 minute 15 minutes before use. Dilute 100×concentrated Biotin-Antibody to a 1×working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent) and use on the same day. 4. Preparation of Enzyme Conjugate Working Solution: 15 minutes before use, centrifuge the 100x concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL concentrate + 990 μL universal diluent). Use the same day. 5. Preparation of 1× Wash Solution: Add 10 mL of 20× Wash Solution to 190 mL of distilled water. (Concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing.) Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return them to 4°C. 2. Sample Addition: Add 50 μL of sample or standard of varying concentrations to the corresponding wells. Add 50 μL of universal diluent to the blank wells, followed by 50 μL of Biotin-Antibody Working Solution to each well. Cover with a film sealer and incubate at 37°C for 1 hour. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the plate to minimize matrix effects. The sample concentration should be multiplied by the dilution factor when calculating the final sample concentration. It is recommended to run replicates for all samples and standards.) 3. Plate Wash: Discard the remaining liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, then shake off the solution and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 4. Enzyme Conjugate Working Solution: Add 100 μL of enzyme conjugate working solution to each well. Cover with a film sealer and incubate at 37°C for 30 minutes. 5. Washing: Discard the liquid and wash the plate five times as in step 3. 6. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with sealing film, and incubate at 37°C in the dark for 15 minutes. 7. Adding stop solution: Remove the plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot a four-parameter logistic function standard curve on double-logarithmic graph paper with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample and retest. Multiply the sample concentration by the corresponding dilution factor.

Sensitivity: 7.2 pg/mL
Specificity: No significant cross-reactivity with other analogues.

Theory

This kit uses a competitive enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled antibody, and HRP conjugate are sequentially added to microwells pre-coated with a universal dopamine (DA) antigen. After incubation and washing, the assay is developed using the substrate TMB. TMB is converted to blue by peroxidase (HRP) catalysis and to yellow by acid. The intensity of the color is positively correlated with the universal dopamine (DA) content in the sample. The absorbance (OD value) is measured at 450 nm using a microplate reader to calculate the sample concentration.

Source

All

Synonym

General Dopamine

Detection Type

Competition Law

Composition

Name	9 6 T Configuration	Remarks
Pre-coated 96-well enzyme plate	8 holes×12 strips	None
Standard	2 bottles	Dilution according to the instructions
Universal diluent	2×20mL	None
Concentrated Biotin-Antibody (100×)	60uL	Dilution according to the instructions
Concentrated enzyme conjugate (100×)	120uL	Dilution according to the instructions
20× Wash solution	2×10mL	Dilute according to instructions
Substrate (TMB)	10mL	None
Stop solution	6mL	none
Sealing film	4 sheets	None
Instructions	1 serving	None

Background

Dopamine, with the chemical formula C₆H₃(OH)₂-CH₂-CH₂-NH₂, is a brain secretion. Its official chemical name is "4-benzene-1,2-diol," or "DA." Dopamine (DA, short for 3,4-dihydroxyphenylethylamine) is a neurotransmitter that plays several important roles in the brain and body. It is an organic chemical in the catecholamine and phenylethylamine families. Dopamine accounts for approximately 80% of the catecholamine content in the brain. It is an amine synthesized by removing a carboxyl group from its precursor chemical, L-DOPA, which is synthesized in the brain and kidneys. Dopamine is also synthesized in plants and most animals. In the brain, dopamine functions as a neurotransmitter—a chemical released by neurons (nerve cells) to signal other nerve cells. Neurotransmitters are synthesized in specific areas of the brain but have systemic effects on many other regions. The brain contains several distinct dopamine pathways, one of which plays a primary role in the motivational component of reward-motivated behavior. The anticipation of most types of rewards increases dopamine levels in the brain, and many addictive drugs either increase dopamine release or block its reuptake into neurons after release. Other brain dopamine pathways are involved in motor control and the release of various hormones. These pathways and cell groups form a dopamine system with neuromodulatory effects. Outside the central nervous system, dopamine primarily acts as a local paracrine messenger. In the blood vessels, it inhibits norepinephrine release and acts as a vasodilator (at normal concentrations); in the kidneys, it increases sodium excretion and urine volume; in the pancreas, it reduces insulin secretion; in the digestive system, it reduces gastrointestinal motility and protects the intestinal mucosa; and in the immune system, it reduces lymphocyte activity. With the exception of the blood vessels, dopamine in each of these peripheral systems is synthesized locally and exerts its effects near the cells where it is released.

General Notes

1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use.
2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation.
3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value.
4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue.
5. Avoid cross-contamination of reagents and specimens to prevent erroneous results.
6. Avoid direct exposure to strong light during storage and incubation.
7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit.
8. Do not use expired products, and do not mix components with different product numbers and batches.
9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized.
10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures.

Storage Temp.

If the unopened kit is stored at 4°C, the shelf life is 6 months.

Test Range

15.6-1000 pg/mL

Applications

Serum, plasma, and other biological fluids

AntBio is a biotechnology group company dedicated to serving life sciences, aiming to help scientists accelerate research and improve work efficiency. AntBio provides comprehensive and high-quality reagent tools for basic research, drug development, and diagnosis, including research grade antibodies, proteins, biochemical reagents, and assay kits. These research tools are widely used in different segments of life science research. The group company currently consists of three brands, Absin, Starter-Bio and UA-Bio.