DESCRIPTION

GRIN1 encodes subunit 1 of the N-methyl-D-aspartate (NMDA) receptor, which is a heteromeric glutamate-gated calcium ion channel essential for synaptic function in the brain. NMDARs play important roles in normal brain development and function, such as synaptic plasticity, neural development, learning and memory. NMDAR dysfunction has been associated with several neurological disorders including Parkinson, Alzheimer and Huntington diseases. Disrupted motor learning and long-term synaptic plasticity in mice lacking NMDAR1 in the striatum.

PRINCIPLE OF THE ASSAY

Sandwich ELISA structure (Detection antibody labeled with HRP) A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody labeled with HRP also binds to the analyte. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns solution yellow. The color intensity is proportional to the quantity of bound protein which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 8 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 16 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	8	2,358.6	128.9	5.5
2	8	567.1	26.3	4.6
3	8	282.4	9.4	3.3

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,507.5	182.0	7.3
2	16	622.6	62.7	10.1
3	16	304.7	30.2	9.9

RECOVERY

The recovery of human/mouse NMDAR1/GRIN1 spiked to three different levels throughout the range of the assay in tissue lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Tissue lysate	1:16	90	70-121

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay.

		Tissue lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	82
	Range (%)	75-89
1:8	Average% of Expected	101
	Range (%)	100-102
1:16	Average% of Expected	117
	Range (%)	114-121

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human/Mouse NMDAR1/GRIN1 Sandwich ELISA Kit produced at Sunlong Medical™.