High Throughput ProcartaPlex 384-well Assays | Thermo Fisher Scientific

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The ProcartaPlex 384-well assays enable multiplex protein analysis in a high throughput format with optimized reagent formulation specifically developed for smaller reaction volumes in 384-well flat bottom plates that require less sample input compared to the 96-well format. This high throughput, magnetic bead-based immunoassay allows for testing of up to 65 analytes per well and can be read on Luminex xMAP instruments—FLEXMAP 3D and INTELLIFLEX.

Comparison of specifications: 96-well vs 384-well ProcartaPlex Assays

	96-well	384-well
Intra-Assay (%CV)	≤15%	≤15%
Inter-Assay (%CV)	≤15%	≤15%
Linearity	4–5 logs	4–5 logs
Plex level	up to 65	up to 65
Sample input	Serum, Plasma, CCS, CSF	Serum, Plasma, CCS, CSF
Compatible Luminex instruments	MAGPIX, Luminex 200, FLEXMAP 3D, INTELLIFLEX	FLEXMAP 3D, INTELLIFLEX
Sample volume	25 µl	6.3 µl

Table 1. Comparison of 96-well and 384-well ProcartaPlex Assays.

ProcartaPlex 384-well formats

The ProcartaPlex 384-well assays are available as exclusive pre-dispensed capture beads on the plate and as liquid formulation of premixed bead sets in a single vial.

ProcartaPlex 384-well (Lyophilized Beads)

ProcartaPlex 384-well assay with Lyophilized bead option

We offer a unique solution with pre-dispensed, lyophilized beads on 384-well flat bottom plates available for Human and Mouse. This convenient solution offers less hands-on time, less time-to-result, and seamless integration into workflows compared to other Luminex multiplex immunoassay offerings.

Customize your panel using the ProcartaPlex Panel Configurator below and select "Lyophilized on plate" for bead formulation or select one of our preconfigured lyophilized panels.

ProcartaPlex Panel Configurator

General workflow- lyophilized bead option

Figure 1. The Invitrogen ProcartaPlex 384-well Multiplex Immunoassay with lyophilized beads eliminates the bead dispensation step in the general workflow minimizing bead dispensation errors.

ProcartaPlex 384-well (Liquid Beads)

ProcartaPlex 384-well assay with liquid bead option

Choose from the largest selection of analytes available in 384-well format for Human, Mouse, Rat, Canine, and NHP. Premixed bead sets are provided as classical liquid formulation provided in a single vial to be pipetted on 384-well flat bottom plates.

Create your unique ProcartaPlex Mix & Match Panel using the ProcartaPlex Panel Configurator below and select “Liquid” for bead formulation.

ProcartaPlex Panel Configurator

Figure 2. General workflow for Invitrogen ProcartaPlex 384-well Multiplex Immunoassay with liquid bead option.

ProcartaPlex 384-well performance characteristics

All ProcartaPlex assays are extensively tested for specificity, sensitivity, and precision to ensure optimal performance. ProcartaPlex 384-well format helps to increase sample throughput while maintaining the robustness of a 96-well assay.

The following data will highlight the differences between the 96-well and 384-well protocols and compare performance data generated using a FLEXMAP3D instrument operated with increased signal enhancing by the photomultiplier tube and an automated washer.

Sensitivity

The ProcartaPlex 384-well format provides increased sample throughput with comparable sensitivity to the 96-well format.

		Sensitivity [pg/ml]
Analyte	Assay range (pg/ml)	96-well	384-well (liquid)	384-well (lyophilized)
Eotaxin (CCL11)	2900—6.3	0.6	0.2	0.5
GRO alpha (CXCL1)	12500—3.1	1.2	0.9	0.7
IL-15	14900—3.6	1.2	1.1	0.9
IL-1 alpha	2550—0.6	0.1	0.1	0.1
IL-1RA	149100-145.6	98.0	49.5	61.5
IL-31	40300—9.8	3.0	5.5	4.9
IL-7	4450—1.1	0.2	0.1	0.1
IL-8 (CXCL8)	14800—3.6	0.9	1.0	0.9
IFN alpha	2050—0.5	0.2	0.2	0.2
IP-10	8700—2.1	0.4	0.5	0.4
MCP-1 (CCL2)	14800—3.6	1.0	0.5	1.6
MIP-1 alpha (CCL3)	15100—3.7	0.5	0.4	0.3
MIP-1 beta (CCL4)	47400—11.6	1.0	0.8	0.7
RANTES (CCL5)	5600—1.4	0.3	0.2	0.2
SDF-1 alpha	73500—17.9	4.2	2.5	3.6
TNF beta	25900—6.3	5.9	3.9	3.9

Table 2. Comparison of sensitivities of analytes from the same standard mix lot in different product formats.

Standard curves

The graphs below compare 7-point standard curves for selected analytes in 96-well and 384-well formats. Similar standard curve performances in 96-well and 384-well formats ensure consistent results regardless of ProcartaPlex assay format.

Sample correlation

A comparison of ProcartaPlex 96-well and 384-well sample values measured on a FLEXMAP 3D instrument for selected analytes is illustrated in the graphs below. Measured sample concentrations for 16 human plasma samples are shown.

The high correlation of sample concentrations across the ProcartaPlex formats ensure reliable and reproducible results.

96- vs 384-well format

The ProcartaPlex 384-well format was specifically developed to increase sample throughput while maintaining the robustness of a 96-well assay. Data of selected analytes in the graphs below illustrate sample correlations between 96-well and 384-well format of 16 human plasma samples.

High sample correlation coefficients (R²) make it easy to switch between 96-well and 384-well formats with consistent results.

MIP-1alpha Plate Format Correlation Data

Lyophilized vs Liquid 384-well format

ProcartaPlex 384-well assays are available in two formats: ready-to-use plates with pre-dispensed lyophilized beads for workflow convenience (384-well lyophilized) and as liquid formulation of bead sets in a vial to be pipetted onto the 384-well flat bottom plate (384-well liquid).

Data of selected analytes in the graphs below illustrate sample correlations between the 384-well lyophilized and 384-well liquid format obtained for 16 human plasma samples.

The high sample coefficients (R²) clearly demonstrate identical performance of ProcartaPlex 384-well assays with lyophilized and liquid bead formulation.