DESCRIPTION

CD80, also known as B7-1, is a type I membrane protein and a member of the immunoglobulin superfamily. It plays a crucial role in T cell activation and regulation by providing a co-stimulatory signal necessary for T cell proliferation and cytokine production. CD80 interacts with CD28 on T cells to promote T cell activation and with CTLA-4 to inhibit T cell activation. This dual role highlights its importance in modulating immune responses. Additionally, CD80 acts as a receptor for adenovirus subgroup B, indicating its involvement in viral infections.

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	184.8	5.9	3.2
2	8	49.6	3.7	7.4
3	8	29.4	2.4	8.1

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	188.0	10.0	5.3
2	16	51.1	4.3	8.4
3	16	28.6	2.6	9.1

RECOVERY

The recovery of mouse CD80/B7-1 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type		Average% of Expected	Range (%)
Mouse serum	1:16	100	98-102
	1:32	103	97-110
Cell lysate	1:256	106	99-110
	1:512	117	111-123

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 mouse serum was initially diluted 1:8. The cell lysate was initially diluted 1:128.)

		Mouse serum	Cell lysate
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	114	99
	Range (%)	107-120	96-102
1:8	Average% of Expected	113	98
	Range (%)	98-127	95-101
1:16	Average% of Expected	103	97
	Range (%)	78-119	80-107

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human Mouse CD80/B7-1 Sandwich ELISA Kit produced at Sunlong Medical™.