"GRIN1" Related Products


Human GRIN1/GRIN2D Stable Cell Line-HEK293

Cat.No.: CSC-RI0177
Cell Line Description: HEK293-HuGRIN1/GRIN2D cell line is a hypotriploid human cell line, which has been co-transfected with a human glutamate receptor, ionotropic, N-methyl D-aspartate 1 (GRIN1) and a human glutamate receptor, ionotropic, N-methyl D-aspartate 2D (GRIN2D) to allow stably express of the human GRIN1 and GRIN2D. It is an example of a cell line transfected using our proprietary CBTGS gene screening and amplification system.
Background: The human GRIN1/GRIN2D genes encode an N-methyl-D-aspartate receptor belonging to the ionotropic glutamate receptor family. NMDA receptors expressed in neurons are therapeutic targets in treatment of neurological disorders.
Growth Properties: Adherent
Morphology: Epithelial
Host Cell: HEK293
Cell Line Validation: 1. Gene expression: qPCR experiments determined specific expression of human GRIN1/GRIN2D.2. Protein expression: GRIN1/GRIN2D expression in this cell line has been validated by WB.3. Functional validation: FLIPR.
Application: Therapeutic Area: neurodegeneration & stroke
Sub-type: Ligand-gated cation channels
Propagation: Complete growth medium: DMEM/Complete growth medium: Ham"s F12; 10% FBS; 500 μg/mL G418Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C
Starting Cells From Frozen Cell Stock: 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed. Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.2. Clean the outside of the vial with 70% ethanol before opening.3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media. 4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.6. Place the cells in the 37°C incubator with 5% CO2.7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
Subculturing: 1. Remove and discard culture medium.2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.5. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:10 is recommended Medium Renewal: 2 to 3 times per week
Mycoplasma: Mycoplasma Status: Negative (MycoAlert Kit)
Freeze Medium: Complete growth medium 90%; DMSO, 10%
Storage: Liquid nitrogen
Preservation: 1. Detach cells from culture dish according to the Sub-Culture Procedure.2. Resuspend cells at a density of 5 x 10^6 cells/mL in freeze medium.Note: A T-75 culture flask typically yields enough cells for preparing two frozen vials.3. Aliquot 1 mL cells into cryogenic vials.4. Place vials in a freezing container and store at –80 °C overnight.5. Transfer vials to liquid nitrogen for long term storage. If properly stored, cells should remain stable for years.
Safety Considerations: The following safety precautions should be observed.1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.2. No eating, drinking or smoking while handling the stable line.3. Wash hands after handling the stable line and before leaving the lab.4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.5. All waste should be considered hazardous.6. Dispose of all liquid waste after each experiment and treat with bleach.
Ship: Dry ice
Gene Name: GRIN2D glutamate receptor, ionotropic, N-methyl D-aspartate 2D [ Homo sapiens ]
Official Symbol: GRIN2D
Synonyms: GRIN2D; glutamate receptor, ionotropic, N-methyl D-aspartate 2D; NMDAR2D; glutamate [NMDA] receptor subunit epsilon-4; EB11; GluN2D; N methyl d aspartate receptor subunit 2D; NR2D; estrogen receptor binding CpG island; N-methyl D-aspartate receptor subtype 2D; N-methyl-d-aspartate receptor subunit 2D;
Gene ID: 2906
mRNA Refseq: NM_000836
Protein Refseq: NP_000827
MIM: 602717
UniProt ID: O15399
Chromosome Location: 19q13.33
Pathway: Activation of NMDA receptor upon glutamate binding and postsynaptic events, organism-specific biosystem; Alcoholism, organism-specific biosystem; Alcoholism, conserved biosystem; Alzheimers disease, organism-specific biosystem; Alzheimers disease, conserved biosystem; Amphetamine addiction, organism-specific biosystem; Amphetamine addiction, conserved biosystem;
Function: N-methyl-D-aspartate selective glutamate receptor activity; extracellular-glutamate-gated ion channel activity; glutamate binding; neurotransmitter binding; receptor activity; transporter activity; voltage-gated cation channel activity;

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