DESCRIPTION

RNASET2 (Ribonuclease T2) is a highly conserved enzyme in a wide range of organisms and belongs to the RNase T2 family. The main biological role of intracellular RNASET2 is to cleave or degrade exogenous or endogenous RNA substrates as well as contributing to mitochondrial RNA metabolism, and also activating immune responses. In addition, it is involved in the regulation of angiogenesis, apoptosis, cell proliferation/differentiation, and immune regulation. In cancer, dysregulated expression of RNASET2 is associated with a variety of pathological conditions, including autoimmune diseases and cancer. The calculated molecular weight of human RNASET2 is approximately 35 kDa, but due to post-translational modifications such as glycosylation, its actual molecular weight may be observed as a mildly glycosylated form with a molecular weight of 38-45 kDa and a highly glycosylated form with a molecular weight of 50-80 kDa.

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 (ng/mL)	SD	CV%
1	8	4.84	0.08	1.65
2	8	1.32	0.04	3.03
3	8	0.43	0.02	4.65

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	4.82	0.10	2.07
2	16	1.31	0.04	3.05
3	16	0.42	0.02	4.76

RECOVERY

The recovery of human RNASET2 spiked to three different levels throughout the range of the assay in human plasma was evaluated.

Sample Type		Average% of Expected	Range (%)
Human plasma	1:8	94	85-101
	1:16	104	101-108

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.

		Human plasma
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	101
	Range (%)	95-107
1:8	Average% of Expected	104
	Range (%)	104-105
1:16	Average% of Expected	116
	Range (%)	111-120

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human RNASET2 produced at Sunlong Medical™.