Rat GRIA1(Glutamate receptor 1)ELISA Kit (RTES01082)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- GLUH1, GLUR1, GLURA, HBGR1, MGC133252, AMPA 1
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||31.25-2000 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Rat GRIA1 in samples. No significant cross-reactivity or interference between Rat GRIA1 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 Rat GRIA1. 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 Rat GRIA1 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 Rat GRIA1, 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 Rat GRIA1. The concentration of Rat GRIA1 in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||GluR1: an integral membrane protein belonging to the glutamate-gated ion channel family. L-glutamate (Glu) acts as an excitatory neurotransmitter at many synapses in the central nervous system. Glutamate receptors are heteromeric protein complexes with multiple subunits, each possessing transmembrane regions, and all arranged to form a ligand-gated ion channel. The postsynaptic actions of Glu are mediated by a variety of receptors that are named according to their selective agonists. Each of the four GluR proteins (GRIA1-4) include flip and flop isoforms generated by alternative RNA splicing. Expressed in granule and pyramidal cells in the hippocampal formation.|
|UniProt Protein Details:|
Protein type:Membrane protein, multi-pass; Channel, calcium; Channel, ligand-gated; Membrane protein, integral
Cellular Component: asymmetric synapse; cell junction; cell soma; cell surface; cytosol; dendrite; dendritic shaft; dendritic spine; early endosome; endoplasmic reticulum; endoplasmic reticulum membrane; excitatory synapse; integral to membrane; ionotropic glutamate receptor complex; membrane; neuromuscular junction; neuron projection; plasma membrane; postsynaptic density; postsynaptic membrane; protein complex; recycling endosome; synapse; synaptic vesicle
Molecular Function:adenylate cyclase binding; alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate selective glutamate receptor activity; beta-2 adrenergic receptor binding; extracellular-glutamate-gated ion channel activity; G-protein alpha-subunit binding; G-protein beta-subunit binding; identical protein binding; ionotropic glutamate receptor activity; myosin V binding; PDZ domain binding; protein binding; protein domain specific binding; protein homodimerization activity; protein kinase A binding; protein kinase binding; small GTPase binding
Biological Process: generation of action potential; ionotropic glutamate receptor signaling pathway; long-term memory; positive regulation of membrane potential; positive regulation of synaptic transmission; receptor internalization; regulation of receptor recycling; regulation of synaptic plasticity; regulation of synaptic transmission; response to arsenic; response to cocaine; response to drug; response to electrical stimulus; response to estradiol stimulus; response to lithium ion; response to organic cyclic substance; response to peptide hormone stimulus; response to toxin; spinal cord development; synaptic transmission; synaptic transmission, glutamatergic
|NCBI Summary:||mouse homolog may play a role in regulation of neuronal synaptic plasticity and conditioned reinforcement learning [RGD, Feb 2006]|
|NCBI GenInfo Identifier:||29789269|
|NCBI Gene ID:||50592|
|NCBI Accession:||NP_113796. 1|
|UniProt Related Accession:||P19490|
|Molecular Weight:||101,555 Da|
|NCBI Full Name:||glutamate receptor 1|
|NCBI Synonym Full Names:||glutamate ionotropic receptor AMPA type subunit 1|
|NCBI Official Symbol:||Gria1|
|NCBI Official Synonym Symbols:||GluA1; gluR-A|
|NCBI Protein Information:||glutamate receptor 1|
|UniProt Protein Name:||Glutamate receptor 1|
|UniProt Synonym Protein Names:||AMPA-selective glutamate receptor 1; GluR-A; GluR-K1; Glutamate receptor ionotropic, AMPA 1; GluA1|
|Protein Family:||Glutamate mutase|
|UniProt Gene Name:||Gria1|
|UniProt Entry Name:||GRIA1_RAT|
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.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat GRIA1 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat GRIA1 were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.25||5.65||5.01||5.45||4.28||5.42|
The recovery of Rat GRIA1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||89-104||96|
|Cell culture media (n=5)||93-107||101|
Samples were spiked with high concentrations of Rat GRIA1 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)|
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.
|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.