Human Signal Transduction ELISA Kits
Human GRIN1 (Glutamate Receptor, Ionotropic, N-Methyl-D-Aspartate 1) ELISA Kit (HUES02634)
- SKU:
- HUES02634
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q05586
- Sensitivity:
- 0.09ng/mL
- Range:
- 0.16-10ng/mL
- ELISA Type:
- Sandwich
- Synonyms:
- GRIN1, GluN1, MRD8, NMDA1, NMDAR1, NR1,Glutamate [NMDA] receptor subunit zeta-1
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Signal Transduction
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 0.16-10 ng/mL |
| Sensitivity: | 0.10 ng/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human GRIN1 in samples. No significant cross-reactivity or interference between Human GRIN1 and analogues was observed. |
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human GRIN1. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human GRIN1 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 GRIN1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human GRIN1. The concentration of Human GRIN1 in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | NMDAR1: a subunit of N-methyl-D-aspartate (NMDA) receptors, members of the glutamate receptor channel superfamily. Possesses high calcium permeability and voltage-dependent sensitivity to magnesium and is modulated by glycine. Plays a key role in synaptic plasticity, synaptogenesis, excitotoxicity, memory acquisition and learning. Mediates neuronal functions in glutamate neurotransmission. Three alternatively-spliced isoforms have been described. |
| UniProt Protein Details: | Protein type:Channel, ligand-gated; Membrane protein, multi-pass; Membrane protein, integral; Channel, calcium Chromosomal Location of Human Ortholog: 9q34. 3 Cellular Component: neuron projection; cell surface; endoplasmic reticulum; integral to plasma membrane; postsynaptic density; dendrite; dendritic spine; terminal button; excitatory synapse; N-methyl-D-aspartate selective glutamate receptor complex; postsynaptic membrane; synaptic vesicle; plasma membrane; synapse; cell junction Molecular Function:voltage-gated cation channel activity; neurotransmitter binding; glutamate receptor binding; calcium channel activity; calcium ion binding; calmodulin binding; protein binding; enzyme binding; glutamate binding; extracellular-glutamate-gated ion channel activity; protein heterodimerization activity; N-methyl-D-aspartate selective glutamate receptor activity; glycine binding Biological Process: regulation of long-term neuronal synaptic plasticity; axon guidance; male mating behavior; prepulse inhibition; adult locomotory behavior; positive regulation of apoptosis; regulation of dendrite morphogenesis; rhythmic process; response to morphine; sensory perception of pain; regulation of axonogenesis; calcium ion homeostasis; synaptic transmission; regulation of respiratory gaseous exchange; conditioned taste aversion; ephrin receptor signaling pathway; visual learning; negative regulation of neuron apoptosis; protein tetramerization; cation transport; synaptic transmission, glutamatergic; response to amphetamine; social behavior; respiratory gaseous exchange; pons maturation; cellular calcium ion homeostasis; regulation of membrane potential; response to ethanol; regulation of synaptogenesis; olfactory learning; long-term memory; suckling behavior; propylene metabolic process; ionotropic glutamate receptor signaling pathway; positive regulation of transcription from RNA polymerase II promoter; cerebral cortex development; regulation of excitatory postsynaptic membrane potential; response to calcium ion Disease: Mental Retardation, Autosomal Dominant 8 |
| NCBI Summary: | The protein encoded by this gene is a critical subunit of N-methyl-D-aspartate receptors, members of the glutamate receptor channel superfamily which are heteromeric protein complexes with multiple subunits arranged to form a ligand-gated ion channel. These subunits play a key role in the plasticity of synapses, which is believed to underlie memory and learning. Cell-specific factors are thought to control expression of different isoforms, possibly contributing to the functional diversity of the subunits. Alternatively spliced transcript variants have been described. [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q05586 |
| NCBI GenInfo Identifier: | 548377 |
| NCBI Gene ID: | 2902 |
| NCBI Accession: | Q05586. 1 |
| UniProt Secondary Accession: | Q05586,P35437, Q12867, Q12868, Q5VSF3, Q5VSF4, Q5VSF5 Q5VSF6, Q5VSF7, A6NLK7, A6NLR1, C9K0X1, |
| UniProt Related Accession: | Q05586 |
| Molecular Weight: | 101,851 Da |
| NCBI Full Name: | Glutamate receptor ionotropic, NMDA 1 |
| NCBI Synonym Full Names: | glutamate receptor, ionotropic, N-methyl D-aspartate 1 |
| NCBI Official Symbol: | GRIN1 |
| NCBI Official Synonym Symbols: | NR1; MRD8; GluN1; NMDA1; NMDAR1 |
| NCBI Protein Information: | glutamate receptor ionotropic, NMDA 1; NMD-R1; glutamate [NMDA] receptor subunit zeta 1; glutamate [NMDA] receptor subunit zeta-1; N-methyl-D-aspartate receptor subunit NR1; N-methyl-D-aspartate receptor channel, subunit zeta-1 |
| UniProt Protein Name: | Glutamate receptor ionotropic, NMDA 1 |
| UniProt Synonym Protein Names: | Glutamate [NMDA] receptor subunit zeta-1; N-methyl-D-aspartate receptor subunit NR1; NMD-R1 |
| Protein Family: | Glutamate receptor ionotropic |
| UniProt Gene Name: | GRIN1 |
| UniProt Entry Name: | NMDZ1_HUMAN |
As the OD 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 (ng/mL) | O.D | Average | Corrected |
| 10 | 2.239 2.277 | 2.258 | 2.204 |
| 5 | 1.561 1.577 | 1.569 | 1.515 |
| 2.5 | 0.904 0.87 | 0.887 | 0.833 |
| 1.25 | 0.386 0.41 | 0.398 | 0.344 |
| 0.63 | 0.239 0.217 | 0.228 | 0.174 |
| 0.32 | 0.158 0.136 | 0.147 | 0.093 |
| 0.16 | 0.1 0.106 | 0.103 | 0.049 |
| 0 | 0.044 0.064 | 0.054 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human GRIN1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human GRIN1 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 (ng/mL) | 0.47 | 1.57 | 4.77 | 0.43 | 1.51 | 4.40 |
| Standard deviation | 0.02 | 0.07 | 0.25 | 0.03 | 0.06 | 0.21 |
| C V (%) | 4.26 | 4.46 | 5.24 | 6.98 | 3.97 | 4.77 |
Recovery
The recovery of Human GRIN1 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 | 95 |
| EDTA plasma (n=5) | 90-102 | 96 |
| Cell culture media (n=5) | 94-106 | 99 |
Linearity
Samples were spiked with high concentrations of Human GRIN1 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 (%) | 91-105 | 95-110 | 91-104 |
| Average (%) | 97 | 100 | 96 | |
| 1:4 | Range (%) | 89-100 | 81-92 | 84-97 |
| Average (%) | 95 | 87 | 90 | |
| 1:8 | Range (%) | 88-103 | 85-98 | 83-95 |
| Average (%) | 94 | 92 | 88 | |
| 1:16 | Range (%) | 87-103 | 83-93 | 86-96 |
| Average (%) | 94 | 88 | 91 |
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 ELISA 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 | 1 vial, 10 mL | 4°C(shading light) |
| Stop Solution | 1 vial, 10 mL | 4°C |
| 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 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
