Human Cell Biology ELISA Kits 4
Human D1R/DRD1 (Dopamine Receptor D1) CLIA Kit (HUES00697)
- SKU:
- HUES00697
- Product Type:
- ELISA Kit
- ELISA Type:
- CLIA Kit
- Size:
- 96 Assays
- Sensitivity:
- 37.5pg/mL
- Range:
- 62.5-4000pg/mL
- ELISA Type:
- Sandwich
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cell Biology
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection method: | Chemiluminescence |
| Detection range: | 62.50-4000 pg/mL |
| Sensitivity: | 37.50 pg/mL |
| Sample volume: | 100µL |
| Sample type: | Serum, plasma and other biological fluids |
| Repeatability: | CV < 15% |
| Specificity: | This kit recognizes Human D1R/DRD1 in samples. No significant cross-reactivity or interference between Human D1R/DRD1 and analogues was observed. |
This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Human D1R/DRD1. Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human D1R/DRD1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human D1R/DRD1, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Human D1R/DRD1. The concentration of Human D1R/DRD1 in the samples can be calculated by comparing the RLU of the samples to the standard curve.
| 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 Protein Details: | Protein type:Receptor, GPCR; Membrane protein, multi-pass; GPCR, family 1; Membrane protein, integral Chromosomal Location of Human Ortholog: 5q35. 1 Cellular Component: endoplasmic reticulum membrane; integral to plasma membrane; plasma membrane; nucleus Molecular Function:protein binding; dopamine receptor activity; dopamine binding; dopamine D1 receptor-like receptor activity Biological Process: synaptic transmission, dopaminergic; peristalsis; elevation of cytosolic calcium ion concentration during G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); behavioral fear response; prepulse inhibition; positive regulation of potassium ion transport; generation of action potential; thermoregulation; G-protein signaling, adenylate cyclase activating pathway; vasodilation; positive regulation of synaptic transmission, glutamatergic; adult walking behavior; dopamine metabolic process; synaptogenesis; behavioral response to cocaine; protein import into nucleus; conditioned taste aversion; positive regulation of cAMP biosynthetic process; visual learning; dopamine receptor, adenylate cyclase activating pathway; grooming behavior; response to drug; dopamine transport; dentate gyrus development; cerebral cortex GABAergic interneuron migration; striatum development; maternal behavior; response to amphetamine; adenylate cyclase activation; regulation of dopamine metabolic process; mating behavior; regulation of dopamine uptake; transmission of nerve impulse; astrocyte development; memory; G-protein signaling, coupled to cyclic nucleotide second messenger; habituation; glucose import; operant conditioning; sensitization; positive regulation of release of sequestered calcium ion into cytosol; dopamine receptor, phospholipase C activating pathway; positive regulation of cell migration |
| NCBI Summary: | This gene encodes the D1 subtype of the dopamine receptor. The D1 subtype is the most abundant dopamine receptor in the central nervous system. This G-protein coupled receptor stimulates adenylyl cyclase and activates cyclic AMP-dependent protein kinases. D1 receptors regulate neuronal growth and development, mediate some behavioral responses, and modulate dopamine receptor D2-mediated events. Alternate transcription initiation sites result in two transcript variants of this gene. [provided by RefSeq, Jul 2008] |
| UniProt Code: | P21728 |
| NCBI GenInfo Identifier: | 118228 |
| NCBI Gene ID: | 1812 |
| NCBI Accession: | P21728. 1 |
| UniProt Secondary Accession: | P21728,Q61616, P18901, |
| UniProt Related Accession: | P21728 |
| Molecular Weight: | 75kDa |
| NCBI Full Name: | D(1A) dopamine receptor |
| NCBI Synonym Full Names: | dopamine receptor D1 |
| NCBI Official Symbol: | DRD1 |
| NCBI Official Synonym Symbols: | DADR; DRD1A |
| NCBI Protein Information: | D(1A) dopamine receptor |
| UniProt Protein Name: | D(1A) dopamine receptor |
| UniProt Synonym Protein Names: | Dopamine D1 receptor |
| Protein Family: | D(1A) dopamine receptor |
| UniProt Gene Name: | DRD1 |
| UniProt Entry Name: | DRD1_HUMAN |
As the RLU values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
| Concentration (pg/mL) | RLU | Average | Corrected |
| 4000 | 58037 60125 | 59081 | 59046 |
| 2000 | 26652 29066 | 27859 | 27824 |
| 1000 | 13925 13043 | 13484 | 13449 |
| 500 | 6181 7029 | 6605 | 6570 |
| 250 | 3325 3161 | 3243 | 3208 |
| 125 | 1670 1492 | 1581 | 1546 |
| 62.50 | 738 772 | 755 | 720 |
| 0 | 33 37 | 35 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human D1R/DRD1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human D1R/DRD1 were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 199.76 | 485.03 | 1849.79 | 194.12 | 468.65 | 1670.02 |
| Standard deviation | 17.16 | 56.41 | 183.13 | 15.94 | 41.76 | 169.84 |
| C V (%) | 8.59 | 11.63 | 9.90 | 8.21 | 8.91 | 10.17 |
Recovery
The recovery of Human D1R/DRD1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 89-102 | 94 |
| EDTA plasma (n=5) | 94-107 | 102 |
| Cell culture media (n=5) | 93-110 | 100 |
Linearity
Samples were spiked with high concentrations of Human D1R/DRD1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
| Serum (n=5) | EDTA plasma (n=5) | Cell culture media (n=5) | ||
| 1:2 | Range (%) | 93-108 | 91-107 | 85-97 |
| Average (%) | 101 | 98 | 91 | |
| 1:4 | Range (%) | 102-114 | 101-118 | 88-100 |
| Average (%) | 108 | 109 | 94 | |
| 1:8 | Range (%) | 94-111 | 89-101 | 96-113 |
| Average (%) | 101 | 95 | 104 | |
| 1:16 | Range (%) | 93-109 | 100-116 | 98-112 |
| Average (%) | 100 | 107 | 104 |
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
| Item | Specifications | Storage |
| Micro CLIA Plate(Dismountable) | 8 wells ×12 strips | -20°C, 6 months |
| Reference Standard | 2 vials | |
| Concentrated Biotinylated Detection Ab (100×) | 1 vial, 120 µL | |
| Concentrated HRP Conjugate (100×) | 1 vial, 120 µL | -20°C(shading light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 4°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | |
| HRP Conjugate Diluent | 1 vial, 14 mL | |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | |
| Substrate Reagent A | 1 vial, 5 mL | 4°C (shading light) |
| Substrate Reagent B | 1 vial, 5 mL | 4°C (shading light) |
| Plate Sealer | 5 pieces | |
| Product Description | 1 copy | |
| Certificate of Analysis | 1 copy |
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids. ) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
- Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.
- Determine the RLU value of each well immediately.
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