Bio-Plex Pro™ Rat Cytokine, Chemokine, and Growth Factor Assays

Representative assay working ranges, assay sensitivity, and precision. The LLOQ, ULOQ, LOD, and inter-assay precision %CV are mean data determined from three independent multiplex assays in a serum-based matrix. Intra-assay %CV is derived from one representative assay. LLOQ and ULOQ are defined as the boundary standard curve points in which the performance specifications of individual standard points were met for <20% intra-assay CV, <30% inter-assay CV, and recovery range of 80–120%. Data were generated using the magnetic workflow with the Bio-Plex Pro II wash station.

	Assay Working Ranges, pg/ml		Assay Sensitivity, pg/ml	Assay Precision
Targets	LLOQ	ULOQ	LOD	Intra-assay %CV	Inter-assay %CV
Singleplex Assays
IL-12p40	2	2,015	0.4	3	4
MIP-2	0.3	1,085	0.2	4	5
Multiplex Assays
EPO	32	32,258	8	4	1
G-CSF	1	2,911	0.2	2	1
GM-CSF	6	31,519	0.6	4	1
GRO-KC	2	1,521	0.6	4	2
IFN-gamma	4	23,955	1	5	3
IL-1alpha	2	24,649	1	3	3
IL-1beta	6	31,666	2	4	2
IL-2	8	7,981	3	4	3
IL-4	1	16,209	1	6	3
IL-5	6	2,007	6	4	0.4
IL-6	13	30,766	10	4	3
IL-7	2	32,472	0.4	4	3
IL-10	4	34,385	5	4	6
IL-12p70	4	15,481	0.7	4	3
IL-13	2	15,751	0.9	4	2
IL-17	1	3,079	0.1	3	2
IL-18	4	31,652	4	4	5
M-CSF	2	23,264	0.4	2	3
MIP-1alpha	25	4,325	12	4	7
MIP-3alpha	1	3,032	0.7	4	2
RANTES	8	7,950	3	5	2
TNF-alpha	16	8,207	3	5	7
VEGF	0	3,963	0.3	3	5

Assay Performance Definitions

Assay working range — the range of concentrations within which the assay is precise and accurate. Boundaries of the assay working range are the lower limit of quantitation (LLOQ) and the upper limit of quantitation (ULOQ).

Precision — the coefficient of variation (%CV) at concentrations within the assay working range

Accuracy — ratio of the observed concentration versus the expected concentration of a known amount of spiked analyte within the assay working range

Sensitivity (Limit of Detection, LOD) — the concentration of analyte for which the fluorescence intensity signal is two-fold of the standard deviation above the background signal

Incubation time	3-Plex Panel (171-W4001)
Sample volume	12.5 µl
Accuracy (recovery range)	80–100%
Precision    Intra-assay %CV    Inter-assay %CV	≤10%    ≤15%

Representative assay working ranges, assay sensitivity, and precision. The LLOQ, ULOQ, LOD, and inter-assay precision %CV are mean data determined from three independent multiplex assays in serum-based matrix. Intra-assay %CV is derived from one representative assay. LLOQ and ULOQ are defined as the boundary standard curve points in which the performance specifications of individual standard points were met for 10% intra-assay CV, 15% inter-assay CV, and recovery range of 80–100%. Data were generated using the magnetic workflow with the Bio-Plex Pro II wash station.

TGF-β Assay Specifications
	Assay Working Ranges, pg/ml		Assay Sensitivity, pg/ml	Assay Precision
Targets	LLOQ	ULOQ	LOD	Intra-assay %CV	Inter-assay %CV
Premixed 3-Plex Assay
TGF-β1	1.69	27,616	3.9	4.5	4.9
TGF-β2	14.7	30,080	1.9	6.3	9.1
TGF-β3	2.8	15,031	0.5	6.9	8.2

Assay Performance Definitions
Assay working range – the range of concentrations within which the assay is precise and accurate. Boundaries of the assay working range are the lower limit of quantitation (LLOQ) and the upper limit of quantitation (ULOQ)

Precision – the coefficient of variation (%CV) at concentrations within the assay working range

Accuracy – ratio of the observed concentration versus the expected concentration of a known amount of spiked analyte within the assay working range

Sensitivity (Limit of Detection, LOD) – the concentration of analyte for which the fluorescence intensity signal is 2-fold of the standard deviation above the background signal.