Dopamine ELISA Kit

Catalog No. ABIN1118052

Product Details Dopamine ELISA Kit

Target: Dopamine (DA)

Reactivity: Various Species

Detection Method: Colorimetric

Method Type: Competition ELISA

Detection Range: 12.35 pg/mL - 1000 pg/mL

Minimum Detection Limit: 12.35 pg/mL

Application: ELISA

Purpose: The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of DA in Serum,Plasma and other Biological Fluids

Sample Type: Plasma, Serum

Analytical Method: Quantitative

Specificity:

This assay has high sensitivity and excellent specificity for detection of Dopamine (DA).
No significant cross-reactivity or interference between Dopamine (DA) and analogues was observed.

Cross-Reactivity (Details): No significant cross-reactivity or interference between Dopamine (DA) and analogues was observed.

Sensitivity: 4.71 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Application Notes:

• Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
• The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
• Kits from different batches may be a little different in detection range, sensitivity and color developing time.
• Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
• Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
• There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
• Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
• Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
• Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
• Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 50 μL

Assay Time: 2 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 50μL standard or sample to each well.
   Then add 50μL prepared Detection Reagent A immediately.
   Shake and mix. Incubate 1 hour at 37 °C,
3. Aspirate and wash 3 times,
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
5. Aspirate and wash 5 times,
6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
7. Add 50μL Stop Solution. Read at 450 nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 1,000pg/mL. Please prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 1,000pg/mL, 333.33pg/mL, 111.11pg/mL, 37.04pg/mL, 12.35pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
4. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
5. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
7. Contaminated water or container for reagent preparation will influence the detection result.

Assay Precision:

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Dopamine (DA) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Dopamine (DA) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Handling Advice: The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Storage: 4 °C

Storage Comment:

• For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
• For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
  Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
• For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.

Expiry Date: 6 months

References for Dopamine ELISA Kit (ABIN1118052)

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Chen, Wu, Zhang, Ma, Liu: "Neurotoxicity of Mn3O4 nanoparticles: Apoptosis and dopaminergic neurons damage pathway." in: Ecotoxicology and environmental safety, Vol. 188, pp. 109909, (2020) (PubMed).

Ci, Li, You, Jin, Zhou, Gao, Hoi, Wang, Chang, Yu: "Iron overload induced by IRP2 gene knockout aggravates symptoms of Parkinson's disease." in: Neurochemistry international, Vol. 134, pp. 104657, (2020) (PubMed).

Bai, Ding, Li, Kong, Xin, Yang, Zhang, Rong, Yao, Lu, Ji, Li, Huang: "Polydatin protects SH-SY5Y in models of Parkinson's disease by promoting Atg5-mediated but parkin-independent autophagy." in: Neurochemistry international, Vol. 134, pp. 104671, (2020) (PubMed).

Kabel, Omar, Alhadhrami, Alharthi, Alrobaian et al.: "Linagliptin potentiates the effect of l-dopa on the behavioural, biochemical and immunohistochemical changes in experimentally-induced Parkinsonism: Role of toll-like receptor 4, TGF-β1, NF-κB and ..." in: Physiology & behavior, Vol. 188, pp. 108-118, (2019) (PubMed).

Peng, Xue, Zhou, Zhang, Wang, Liu, Sang, Wang, Tan: "Repetitive transcranial magnetic stimulation inhibits Sirt1/MAO-A signaling in the prefrontal cortex in a rat model of depression and cortex-derived astrocytes." in: Molecular and cellular biochemistry, Vol. 442, Issue 1-2, pp. 59-72, (2018) (PubMed).

Sharma, Taliyan: "High fat diet feeding induced insulin resistance exacerbates 6-OHDA mediated neurotoxicity and behavioral abnormalities in rats." in: Behavioural brain research, Vol. 351, pp. 17-23, (2018) (PubMed).

Yousef, Khalil, Abdou et al.: "Neuro- and nephroprotective effect of grape seed proanthocyanidin extract against carboplatin and thalidomide through modulation of inflammation, tumor suppressor protein p53, neurotransmitters, ..." in: Toxicology reports, Vol. 5, pp. 568-578, (2018) (PubMed).

Mohamed, Abd El-Moneim: "Ginkgo biloba extract alleviates oxidative stress and some neurotransmitters changes induced by aluminum chloride in rats." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 35, pp. 93-99, (2017) (PubMed).

Kandil, Abdelkader, El-Sayeh, Saleh: "Imipramine and amitriptyline ameliorate the rotenone model of Parkinson's disease in rats." in: Neuroscience, Vol. 332, pp. 26-37, (2016) (PubMed).

Ibrahim: "Neuroprotective effect of Cucumis melo Var. flexuosus leaf extract on the brains of rats with streptozotocin-induced diabetes." in: Metabolic brain disease, Vol. 32, Issue 1, pp. 69-75, (2016) (PubMed).

Abdelkader, Safar, Salem: "Ursodeoxycholic Acid Ameliorates Apoptotic Cascade in the Rotenone Model of Parkinson's Disease: Modulation of Mitochondrial Perturbations." in: Molecular neurobiology, (2015) (PubMed).

Safar, Arab, Rizk, El-Maraghy: "Bone Marrow-Derived Endothelial Progenitor Cells Protect Against Scopolamine-Induced Alzheimer-Like Pathological Aberrations." in: Molecular neurobiology, (2015) (PubMed).

Abdel-Ghany, Nabil, Abdel-Aal, Barakat: "Nalbuphine could decrease the rewarding effect induced by tramadol in mice while enhancing its antinociceptive activity." in: European journal of pharmacology, Vol. 758, pp. 11-5, (2015) (PubMed).

Abdelsalam, Safar: "Neuroprotective effects of vildagliptin in rat rotenone Parkinson's disease model: role of RAGE-NF?B and Nrf2-antioxidant signaling pathways." in: Journal of neurochemistry, Vol. 133, Issue 5, pp. 700-7, (2015) (PubMed).

Wang, Wang, Chu, Mu, Wang, Liu, Xie, Yang, Dong, Yuan: "Effect of salt intake and potassium supplementation on urinary renalase and serum dopamine levels in Chinese adults." in: Cardiology, Vol. 130, Issue 4, pp. 242-8, (2015) (PubMed).

Kotb El-Sayed, Amin: "Catha edulis chewing effects on treatment of paranoid schizophrenic patients." in: Neuropsychiatric disease and treatment, Vol. 11, pp. 1067-76, (2015) (PubMed).

Zbroch, Koc-Zorawska, Malyszko, Malyszko, Mysliwiec: "Circulating levels of renalase, norepinephrine, and dopamine in dialysis patients." in: Renal failure, Vol. 35, Issue 5, pp. 673-9, (2013) (PubMed).

Target Details for Dopamine

Target: Dopamine (DA)

Alternative Name: Dopamine (DA Products)

Target Type: Chemical