DESCRIPTION

CSF2RA, also known as CD116, is the alpha subunit of the heterodimeric receptor for colony-stimulating factor 2 (GM-CSF). CSF2RA plays a crucial role in the proliferation, differentiation, and functional activation of hematopoietic cells. CSF2RA is overexpressed in certain cancers and most immune cells, which may contribute to the activation of the immune system. The CSF2RA deletion in mice has been shown to attenuate the pro-inflammatory response and improve survival in the context of acute lung injury (ALI) induced by ricin toxin (RT).

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	1,350.8	87.2	6.5
2	8	323.9	19.1	5.9
3	8	154.1	8.9	5.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	1,350.2	88.4	6.5
2	16	324.5	15.8	4.9
3	16	156.3	9.0	5.7

RECOVERY

The recovery of human AFP spiked to three different levels throughout the range of the assay was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:64	90	83-96
Cell lysate	1:320	86	82-89

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 human plasma was initially diluted 1:4. The cell lysate was initially diluted 1:20. )

		Human plasma	Cell lysate
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	105	101
	Range (%)	99-110	90-111
1:8	Average% of Expected	109	107
	Range (%)	104-115	93-122
1:16	Average% of Expected	106	120
	Range (%)	102-109	118-122

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human CSF2RA/CD116 One-Step ELISA Kit produced at Sunlong Medical™.