DESCRIPTION

TRIM27 (Tripartite Motif-Containing Protein 27), also known as RFP (Ret Finger Protein), is a member of the TRIM family characterized by its conserved RING finger, B-box, and coiled-coil domains. Functioning primarily as an E3 ubiquitin ligase, TRIM27 regulates protein stability and degradation through the ubiquitin-proteasome system, impacting diverse cellular processes such as immune response, transcriptional regulation, and viral defense. It plays a dual role in cancer, acting as either an oncogene or tumor suppressor depending on the cellular context, and is implicated in diseases like autoimmune disorders and viral infections. Its involvement in NF-κB and interferon signaling further highlights its importance in immune modulation, making TRIM27 a potential therapeutic target in oncology and immunology research.

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 (ng/mL)	SD	CV%
1	8	10.96	0.33	3.06
2	8	2.81	0.07	2.64
3	8	1.45	0.05	3.22

Inter-assay Precision

Sample	n	Mean (ng/mL)	SD	CV%
1	16	10.75	0.37	3.47
2	16	2.76	0.11	4.06
3	16	1.45	0.04	2.75

RECOVERY

The recovery of human TRIM27 spiked to three different levels throughout the range of the assay in cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:32	86	81-93
	1:64	84	77-97

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 1:4.)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	96
	Range (%)	86-105
1:8	Average% of Expected	89
	Range (%)	86-92
1:16	Average% of Expected	121
	Range (%)	115-127

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Human TRIM27 Sandwich ELISA Kit produced at Sunlong Medical™.