Mouse D(2) dopamine receptor (Drd2) ELISA Kit

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SKU:
MOEB0527
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P61168
Range:
0.312-20 ng/mL
ELISA Type:
Sandwich
Synonyms:
Drd2, D2R, DRD2, Dopamine Receptor D2, Dopamine D2 receptor
Reactivity:
Mouse
Frequently bought together:

Description

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Product Name:Mouse D(2) dopamine receptor (Drd2) ELISA Kit
Product Code:MOEB0527
Alias:D(2) dopamine receptor, Dopamine D2 receptor, Drd2
Uniprot:P61168
Reactivity:Mouse
Range:0.312-20 ng/mL
Detection Method:Sandwich
Size:96 Assay
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:DRD2: Dopamine receptor whose activity is mediated by G proteins which inhibit adenylyl cyclase. Defects in DRD2 are associated with dystonia type 11 (DYT11); also known as alcohol-responsive dystonia. DYT11 is a myoclonic dystonia. Dystonia is defined by the presence of sustained involuntary muscle contractions, often leading to abnormal postures. DYT11 is characterized by involuntary lightning jerks and dystonic movements and postures alleviated by alcohol. Inheritance is autosomal dominant. The age of onset, pattern of body involvement, presence of myoclonus and response to alcohol are all variable. Belongs to the G-protein coupled receptor 1 family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Receptor, GPCR; GPCR, family 1; Membrane protein, multi-pass; Membrane protein, integral

Cellular Component: synaptic vesicle membrane; integral to plasma membrane; dendrite; postsynaptic density; dendritic spine; integral to membrane; acrosome; perikaryon; cytosol; membrane; endocytic vesicle; axon; plasma membrane; cytoplasmic vesicle; nerve terminal; lateral plasma membrane

Molecular Function:G-protein coupled receptor activity; dopamine D2 receptor-like receptor activity; identical protein binding; ionotropic glutamate receptor binding; signal transducer activity; protein homodimerization activity; dopamine receptor activity; protein heterodimerization activity; dopamine binding; drug binding; receptor binding

Biological Process: elevation of cytosolic calcium ion concentration during G-protein signaling, coupled to IP3 second messenger (phospholipase C activating); positive regulation of dopamine uptake; prepulse inhibition; positive regulation of receptor internalization; thermoregulation; positive regulation of multicellular organism growth; regulation of phosphoprotein phosphatase activity; adult walking behavior; dopamine metabolic process; negative regulation of insulin secretion; dopamine receptor, adenylate cyclase inhibiting pathway; protein localization; negative regulation of blood pressure; phosphatidylinositol metabolic process; response to drug; positive regulation of neuroblast proliferation; response to light stimulus; cerebral cortex GABAergic interneuron migration; negative regulation of synaptic transmission, glutamatergic; regulation of sodium ion transport; arachidonic acid secretion; regulation of synaptic transmission, GABAergic; G-protein signaling, coupled to cAMP nucleotide second messenger; positive regulation of growth hormone secretion; acid secretion; G-protein coupled receptor internalization; reduction of cytosolic calcium ion concentration; regulation of MAPKKK cascade; activation of protein kinase activity; dopamine receptor signaling pathway; positive regulation of transcription from RNA polymerase II promoter; regulation of long-term neuronal synaptic plasticity; synaptic transmission, dopaminergic; peristalsis; startle response; branching morphogenesis of a nerve; negative regulation of circadian sleep/wake cycle, sleep; response to morphine; locomotory behavior; behavioral response to ethanol; signal transduction; regulation of dopamine secretion; negative regulation of cell proliferation; synaptic transmission; behavioral response to cocaine; forebrain development; adenohypophysis development; visual learning; nerve-nerve synaptic transmission; feeding behavior; circadian regulation of gene expression; negative regulation of dopamine secretion; negative regulation of cell migration; associative learning; grooming behavior; regulation of synaptic transmission; positive regulation of cytokinesis; negative regulation of protein secretion; Wnt receptor signaling pathway; negative regulation of protein kinase B signaling cascade; regulation of heart rate; regulation of potassium ion transport; regulation of cAMP metabolic process; sensory perception of smell; negative regulation of adenylate cyclase activity; response to amphetamine; regulation of dopamine uptake; auditory behavior; response to cocaine; G-protein coupled receptor protein signaling pathway; regulation of systemic arterial blood pressure by neurological process; negative regulation of dopamine receptor signaling pathway; axonogenesis; pigmentation; long-term memory; positive regulation of G-protein coupled receptor protein signaling pathway; release of sequestered calcium ion into cytosol; adult behavior; response to hypoxia; regulation of synapse structural plasticity; dopamine receptor, phospholipase C activating pathway

UniProt Code:P61168
NCBI GenInfo Identifier:148747212
NCBI Gene ID:13489
NCBI Accession:NP_034207.2
UniProt Secondary Accession:P61168,P13953, Q0VGH9,
UniProt Related Accession:P61168
Molecular Weight:47,625 Da
NCBI Full Name:D(2) dopamine receptor
NCBI Synonym Full Names:dopamine receptor D2
NCBI Official Symbol:Drd2  
NCBI Official Synonym Symbols:D2R; Drd-2  
NCBI Protein Information:D(2) dopamine receptor
UniProt Protein Name:D(2) dopamine receptor
UniProt Synonym Protein Names:Dopamine D2 receptor
Protein Family:D(2) dopamine receptor
UniProt Gene Name:Drd2  
UniProt Entry Name:DRD2_MOUSE
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment 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

*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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.

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