DESCRIPTION

IFNGR1 (Interferon Gamma Receptor 1) is a cytokine critical for innate and adaptive immunity against viral and intracellular bacterial infections and for tumor control. Cellular responses to IFN-γ are activated through its interaction with a heterodimeric receptor consisting of two subunits, IFNGR1 (CD119) and IFNGR2. IFNGR1 is the ligand-binding subunit which binds IFN-γ with high affinity, whereas IFNGR2 serves as the accessory subunit. Two subunits bind one IFN-γ dimer. Defects in IFNGR1 are a cause of mendelian susceptibility to mycobacterial disease (MSMD), also known as familial disseminated atypical mycobacterial infection.

PRINCIPLE OF THE ASSAY

An anti-tag antibody is pre-coated onto the bottom of wells. After adding antigen or samples, Tagged-Capture antibody and HRP-conjugated detection antibody, a sandwich complex is formed in the solution. TMB acts as a HRP substrate, and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns the 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	2,585.6	59.6	2.3
2	8	635.9	10.2	1.6
3	8	313.8	6.3	2.0

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,541.8	96.7	3.8
2	16	626.8	20.7	3.3
3	16	307.8	11.2	3.6

RECOVERY

The recovery of mouse IFNGR1/CD119 spiked to three different levels throughout the range of the assay in lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Lysate	1:64	74	71-79

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. (The cell lysate was initially diluted 1:5.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	85
	Range (%)	84-86
1:8	Average% of Expected	99
	Range (%)	99-100
1:16	Average% of Expected	111
	Range (%)	111-112

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse IFNGR1/CD119 One-Step ELISA Kit produced at Sunlong Medical™.