DESCRIPTION

CCL6, formerly known as C10 or MRP-1 (Macrophage Inflammatory Protein-Related Protein-1), is a small secretory protein belonging to the CC chemokine family. Chemokines are a large family of cytokines, or signaling proteins, renowned for their ability to induce directed migration (chemotaxis) of nearby responsive cells. CCL6 is a crucial CC chemokine in mice that acts primarily through the CCR1 receptor to orchestrate the migration and activation of key immune cells. It is a central mediator in inflammation, host defense, and pathological processes like cancer and fibrosis.

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	2,812.3	79.7	2.8
2	8	571.6	17.0	3.0
3	8	275.0	8.7	3.2

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	16	2,790.0	107.8	3.9
2	16	571.0	17.4	3.0
3	16	269.7	9.2	3.4

RECOVERY

The recovery of mouse CCL6 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	82	70-98
Tissue lysate	1:32	103	102-104

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 tissue lysate was initially diluted 1:2.)

		Mouse serum(Sample Diluent PT 4B1-ef)	Tissue lysate(Sample Diluent PT 4B1-ec)
1:2	Average% of Expected	100	100
	Range (%)	-	-
1:4	Average% of Expected	97	104
	Range (%)	96-98	101-107
1:8	Average% of Expected	95	110
	Range (%)	92-99	107-113
1:16	Average% of Expected	108	101
	Range (%)	107-109	82-120

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant mouse CCL6 produced at Sunlong Medical™.