Human DRD1 / Dopamine Receptor D1 ELISA Kit (HUFI01194)
- SKU:
- HUFI01194
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P21728
- Sensitivity:
- 0.188ng/ml
- Range:
- 0.313-20ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- D1R, DRD1, Dopamine Receptor D1, D, 1A dopamine receptor, Dopamine D1 receptor
- Reactivity:
- Human
- Research Area:
- Cell Biology
Description
Human DRD1 / Dopamine Receptor D1 ELISA Kit
DRD1/Dopamine Receptor D1 binds to dopamine receptors in the brain and is activated by adenylyl cyclase. DRD1/Dopamine Receptor D1 plays a critical role in neuronal growth and development, mediates some behavioral responses, and influences dopamine receptor D2-mediated events. Cerebral Meningioma and Pathological Gambling are two examples of diseases linked to the DRD1 gene. Signaling by GPCR and G alpha signaling events are among the downstream pathways of DRD1/Dopamine Receptor D1.
Key Features
| Save Time | Pre-coated 96 well plate | |
| Quick Start | Kit includes all necessary reagents | |
| Publication Ready | Reproducible and reliable results |
Overview
| Product Name: | Human DRD1 / Dopamine Receptor D1 ELISA Kit |
| Product Code: | HUFI01194 |
| Size: | 96 Assays |
| Alias: | D1R, DRD1, Dopamine Receptor D1, D, 1A dopamine receptor, Dopamine D1 receptor |
| Detection Method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human DRD1 concentrations in serum plasma and other biological fluids. |
| Sensitivity: | 0.188ng/ml |
| Range: | 0.313-20ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
Additional Information
| Recovery: | Matrices listed below were spiked with certain level of Human DRD1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human DRD1 in samples.
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human DRD1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
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| CV(%): | Intra-Assay: CV<8% |
Kit Components
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8x12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/ -20°C |
| Sample/Standard Dlution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody (Concentrated) | 120ul | 4°C (Protection from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate (SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protection from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer (25X) | 30ml | 4°C |
| Plate Sealer | 5 | - |
Other materials required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
Protein Information
| Uniprot: | |
| UniProt Protein Function: | DRD1: a G-protein coupled receptor.One of the five types (D1 to D5) of receptors for dopamine. The most abundant dopamine receptor in the central nervous system. The activity of this receptor is mediated by G proteins which activate adenylyl cyclase. Interacts with calcyon. |
| UniProt Code: | |
| NCBI GenInfo Identifier: | |
| NCBI Gene ID: | |
| NCBI Accession: | |
| UniProt Related Accession: | |
| Molecular Weight: | 75kDa |
Protocol
*Note: Protocols are specific to each batch/lot. For the exact instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Procedure |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37°C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
Sample Type
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
DRD1 Background
The dopamine receptor D1, also known as DRD1, is a subtype of dopamine receptor that plays a crucial role in the regulation of dopamine-mediated signaling in the central nervous system. Dopamine is an important neurotransmitter involved in several physiological functions, including mood regulation, motor control, cognition, and reward processing.
Function of DRD1 Receptor
The main function of the DRD1 receptor is to mediate the excitatory effects of dopamine. When dopamine binds to the DRD1 receptor, it activates the receptor, leading to the activation of stimulatory G proteins (Gs). The activated Gs proteins, in turn, stimulate the production of a molecule called cyclic adenosine monophosphate (cAMP) within the neuron.
Increased levels of cAMP then initiate a cascade of intracellular events, ultimately leading to various physiological responses in the brain. These responses can include enhanced neural activity, increased alertness, and improved cognitive functions.
Distribution of DRD1 Receptor in the Brain
The DRD1 receptor is widely distributed in the brain, with particularly high concentrations found in areas such as the striatum and the prefrontal cortex. These brain regions are involved in motor control, executive functions, and decision-making, which underscores the importance of DRD1 in these cognitive processes.
Implications of DRD1 Dysfunction
Dysfunction or dysregulation of the DRD1 receptor has been associated with various neuropsychiatric disorders, including schizophrenia, bipolar disorder, and attention-deficit/hyperactivity disorder (ADHD). As a result, research on dopamine receptors like DRD1 continues to be a focus in understanding the underlying mechanisms of these disorders and developing potential therapeutic interventions.
DRD1 FAQs
What is the DRD1 ELISA Kit?
The DRD1 ELISA Kit is an advanced assay designed to accurately quantify Human Dopamine Receptor D1 levels in biological samples.
What are the advantages of using the DRD1 ELISA Kit?
The DRD1 ELISA Kit offers high sensitivity, specificity for Human DRD1, an easy-to-use protocol, quantitative results, compatibility with various samples, and time-efficiency for research screening.
Where can I find more information about the - ELISA Kit?
For more detailed information about the DRD1 ELISA Kit, including technical specifications, performance characteristics, and ordering details, please refer to the product brochure or contact our customer support team. We are here to assist you with any inquiries you may have.
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