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Rat Glutamate [NMDA] receptor subunit zeta-1 (Grin1) ELISA Kit (RTEB0227)

SKU:
RTEB0227
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P35439
Range:
0.312-20 ng/mL
ELISA Type:
Sandwich
Synonyms:
GRIN1, Glutamate Receptor, Ionotropic, N-Methyl-D-Aspartate 1, GluN1, MRD8, NMDA1, NMDAR1, NMD-R1
Reactivity:
Rat
€649
Frequently bought together:

Description

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Rat Glutamate [NMDA] receptor subunit zeta-1 (Grin1) ELISA Kit

The Rat Glutamate NMDA Receptor Subunit Zeta-1 (GRIN1) ELISA Kit is specifically designed for the accurate measurement of GRIN1 levels in rat samples, including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures precise and reliable results, making it ideal for a variety of research applications in neuroscience and pharmacology.GRIN1 is a key subunit of the NMDA receptor, which plays a critical role in synaptic plasticity, learning, and memory. Dysregulation of NMDA receptors has been implicated in various neurological disorders, such as Alzheimer's disease, schizophrenia, and epilepsy.

By quantifying GRIN1 levels, researchers can gain valuable insights into the mechanisms underlying these conditions and potentially identify new therapeutic targets.Overall, the Rat GRIN1 ELISA Kit offers a valuable tool for studying NMDA receptor function and exploring its implications in neurological health and disease. Its ease of use and robust performance make it a valuable addition to any laboratory conducting research in this field.

Product Name:Rat Glutamate [NMDA] receptor subunit zeta-1 (Grin1) ELISA Kit
SKU:RTEB0227
Size:96T
Target:Rat Glutamate [NMDA] receptor subunit zeta-1 (Grin1)
Synonyms:Glutamate [NMDA] receptor subunit zeta-1, N-methyl-D-aspartate receptor subunit NR1, NMD-R1, GluN1, Nmdar1
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Rat
Detection Range:0.312-20ng/mL
Sensitivity:0.162ng/mL
Intra CV:4.8%
Inter CV:8.6%
Linearity:
Sample1:21:41:81:16
Serum(N=5)87-96%105-116%105-115%91-104%
EDTA Plasma(N=5)112-122%112-122%83-94%88-98%
Heparin Plasma(N=5)104-113%117-127%99-108%107-118%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8080-85
Plasma8180-87
Function:Component of NMDA receptor complexes that function as heterotetrameric, ligand-gated ion channels with high calcium permeability and voltage-dependent sensitivity to magnesium. Channel activation requires binding of the neurotransmitter glutamate to the epsilon subunit, glycine binding to the zeta subunit, plus membrane depolarization to eliminate channel inhibition by Mg(2+) (PubMed:1350383, PubMed:1834949, PubMed:1388270, PubMed:8428958, PubMed:18177891, PubMed:28384476, PubMed:15996549, PubMed:24876489, PubMed:27135925, PubMed:27618671, PubMed:28468946). Sensitivity to glutamate and channel kinetics depend on the subunit composition (PubMed:28384476).
Uniprot:P35439
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant rat Glutamate receptor ionotropic, NMDA 1
Sub Unit:Heterotetramer. Forms heterotetrameric channels composed of two zeta subunits (GRIN1), and two epsilon subunits (GRIN2A, GRIN2B, GRIN2C or GRIN2D) (in vitro) (PubMed:18177891, PubMed:28384476, PubMed:16281028, PubMed:15996549, PubMed:21389213, PubMed:24876489, PubMed:27135925, PubMed:28468946, Ref.32). Can also form heterotetrameric channels that contain at least one zeta subunit (GRIN1), an epsilon subunit, plus GRIN3A or GRIN3B (in vitro) (PubMed:11160393, PubMed:11929923, PubMed:12391275). In vivo, the subunit composition may vary in function of the expression levels of the different subunits (Probable). Found in a complex with GRIN2A or GRIN2B, GRIN3A and PPP2CB (PubMed:11588171). Found in a complex with GRIN2A or GRIN2B and GRIN3B (By similarity). Interacts with SNX27 (via PDZ domain); the interaction is required for recycling to the plasma membrane when endocytosed and prevent degradation in lysosomes (By similarity). Interacts with DLG4 and MPDZ (PubMed:15312654). Interacts with LRFN1 and LRFN2 (PubMed:16495444, PubMed:16630835). Interacts with MYZAP (PubMed:18849881). Found in a complex with DLG4 and PRR7 (PubMed:27458189). Found in a complex with GRIN2B and PRR7 (PubMed:27458189). Interacts with PRR7; the interaction is reduced following NMDA receptor activity (PubMed:27458189).
Research Area:Neurosciences
Subcellular Location:Cell membrane Multi-pass membrane protein Cell junction Synapse Postsynaptic cell membrane Cell junction Synapse Postsynaptic cell membrane Postsynaptic density Enriched in postsynaptic plasma membrane and postsynaptic densities.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:NMDA receptor subtype of glutamate-gated ion channels possesses high calcium permeability and voltage-dependent sensitivity to magnesium. Mediated by glycine. Plays a key role in synaptic plasticity, synaptogenesis, excitotoxicity, memory acquisition and learning. It mediates neuronal functions in glutamate neurotransmission. Is involved in the cell surface targeting of NMDA receptors.
NCBI Summary:subunit of NMDA-preferring ionotropic glutamate receptors; may play a role in long term potentiation [RGD, Feb 2006]
UniProt Code:P35439
NCBI GenInfo Identifier:396578144
NCBI Gene ID:24408
NCBI Accession:NP_001257531.1
UniProt Secondary Accession:P35439,Q62646,
UniProt Related Accession:P35439
Molecular Weight:101,987 Da
NCBI Full Name:glutamate receptor ionotropic, NMDA 1 isoform 1b
NCBI Synonym Full Names:glutamate ionotropic receptor NMDA type subunit 1
NCBI Official Symbol:Grin1
NCBI Official Synonym Symbols:NR1; GluN1; NMDAR1
NCBI Protein Information:glutamate receptor ionotropic, NMDA 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:Malate dehydrogenase
UniProt Gene Name:Grin1
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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