DESCRIPTION

CCL4L1 encodes a secreted chemokine ligand in the intercrine beta family that functions in inflammatory and immunoregulatory processes. The encoded 92-amino-acid protein is similar to chemokine (C-C motif) ligand 4, and the gene belongs to a cluster of cytokine genes on the q-arm of chromosome 17. CCL4L1 is localized to the extracellular region, placing it in a position to act in intercellular signaling.

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	283.8	4.6	1.6
2	8	68.0	2.0	2.9
3	8	34.1	2.0	5.9

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	294.5	14.2	4.8
2	16	70.2	3.2	4.6
3	16	36.8	3.1	8.4

RECOVERY

The recovery of human CCL4L spiked to three different levels throughout the range of the assay in human serum was evaluated.

Sample Type		Average% of Expected	Range (%)
Human serum	1:8	103	90-114
	1:16	113	112-115

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 serum
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	108
	Range (%)	95-120
1:8	Average% of Expected	109
	Range (%)	100-117
1:16	Average% of Expected	110
	Range (%)	103-117

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human CCL4L produced at Sunlong Medical™.