Gain deep insights with complementary and highly correlative multiplex panels. You can measure gene expression and protein on a single platform enabling you to have confidence in your results. Get more information from your samples by analyzing up to 80 proteins per well—almost twice as many as conventional xMAP assays. It enables efficient immune response profiling and biomarker discovery. Analyze different types of samples including serum and plasma cell lysates, cell culture supernatants or tissue homogenates CCS. Get deep insight in a single assay, helping save time and precious sample which can help reduce consumables cost. ProcartaPlex and QuantiGene preconfigured panels come in a ready-to-use format.
How Luminex xMAP technology supports multi-omics applications
Invitrogen ProcartaPlex and QuantiGene Plex assays enable a unique high-throughput multi-omics approach by utilizing the Luminex xMAP (Multi-Analyte Profiling) technology. Easily combine genomic and proteomic workflows without compromising data interpretation or sensitivity.
Examination of both RNA and protein expression variations can provide a robust and accurate indication of cell states (see workflow and sample data below). RNA levels can be transient and may not correlate with protein levels, which are likely to be maintained for longer periods of time to maintain cellular functions. Due to differences in timing and expression levels, one assay may detect changes that the other does not. Alternatively, the two data sets can further support key findings. The ability to perform both RNA and protein measurements using simple, unified high-throughput workflows can make the Invitrogen ProcartaPlex and QuantiGene Plex assays well suited for tools for large-scale screening studies that benefit from automation workflows.
Analyze both protein and gene expression from a single sample
U937 cells were stimulated with PMA (24 hr) or LPS (48 hr) and the cell culture supernatant was collected to measure secreted proteins using the Th1/Th2 cytokine and chemokine 20-plex ProcartaPlex panel. The cells were lysed using the QuantiGene lysis mixture and the cell lysate was directly used to measure gene expression changes with the 36-plex QuantiGene Plex panel. Both assays were read on a Luminex xMAP INTELLIFLEX DR-SE instrument.
High-Plex multiplexing offers a transformative advantage for biomarker discovery and validation by enabling simultaneous assessment of up to 80 analytes on the Luminex instrument within a single experiment. This approach greatly improves efficiency by helping conserve resources, precious samples and time, while also providing a holistic view of complex biological interactions.
To demonstrate the power of the 80-plex as a screening method, 30 samples were analyzed. 23 human serum samples were untreated, and 7 plasma samples were treated with LPS 100 ng/mL for 20 hours.
These were quantified in parallel using Invitrogen ProcartaPlex Human Immune Response Panel (80-plex). Differences in the expression pattern of 80 targets in untreated human serum samples and pre-treated plasma samples are shown in the following figures, grouped by low, medium, and high expression levels. Analytes are displayed on the x-axis, and mean concentration for untreated (blue lines) versus treated (orange lines) samples for each analyte on the y-axis.
Click image to enlarge
Click image to enlarge
Click image to enlarge
Figure 1. Differences in the expression pattern of 80 targets in untreated human serum samples and pre-treated plasma samples.
Additionally, a few analytes in the medium expressed samples are shown in more detail. These analytes showed the greatest degree of difference between treated versus untreated samples. This data is presented in four figures as demonstrated here.
Click image to enlarge
Figure 2. Concentration levels for a few analytes with medium expression levels.
To confirm scalability of ProcartaPlex multiplex immunoassays, human serum and stimulated plasma samples were run in parallel using both the Invitrogen ProcartaPlex Chemokine Panel 2 (9-plex) and the Invitrogen ProcartaPlex Human Immune Response Panel (80-plex). Experiments demonstrated high consistency between large-scale versus small-scale multiplex panels, verifying that high content screening can be performed without compromising the test results. Concentration levels for MCP-4 and CCL1 at different serum samples for small plex (9-plex) versus 80-plex is presented in the following figure. This was a method to demonstrate the level of correlation between small and big panels.
Click image to enlarge
Figure 3. Concentration levels for MCP-4 and CCL1 at different serum samples for small plex (9-plex) vs 80-plex.
To further demonstrate the degree of correlation between small and big plex, a regression analysis was performed. This yielded a R2 value >0.9. (0.99 for MCP-4, 0.93 for CCL1).
Click image to enlarge
Figure 4. Correlation Analysis between small and big plex.
Targets highlighted in bold are additional in the Immune Response 64-Plex Mouse ProcartaPlex Panel compared to the Immune Monitoring 48-Plex Mouse ProcartaPlex Panel (EPX480-20834-901).
The ability to scale the number of analytes and correlate data in multiplex experiments is critical for the progression of many projects. For example, it is not uncommon to begin a project by analyzing a large, comprehensive panel on few samples to determine which analytes are affected by a particular disease or drug treatment. The resulted list of analytes can then be investigated further with a smaller panel on a larger set of samples. Thus, there is a great need for scalability in multiplex panel assays to correlate data across different stages of investigation.
To demonstrate the scalability and reproducible performance regardless of plex size mouse serum and plasma samples of healthy, untreated mice were run in parallel on the ProcartaPlex Mouse Immune Monitoring Panel 48-Plex and the ProcartaPlex Mouse Immune Response Panel 64-Plex. Data in Figure 5 and 6 illustrates the high correlation for sample measurements for the same analytes in a large and small panel.
Click image to enlarge
Figure 5. Comparison data for selected analytes in blood samples from healthy, untreated mice.
Click image to enlarge
Click image to enlarge
Figure 6. Data illustrate the high correlation between large and small-scale ProcartaPlex panels.
Performance data from the largest High-Plex QuantiGene Plex assays
Immune Response 80-Plex Human QuantiGene Plex Panel
PBMC were stimulation with Lipopolysaccharide (LPS) and the gene expression change over time was measured with the Immune Response 80-Plex Human QuantiGene Plex. The majority of the 80 gene targets tested showed expression changes over unstimulated control samples with 10 µg/mL LPS-stimulation of PBMCs at 3 time points (see figure 7). Raw MFI data from the QuantiGene Plex assay using the Immune Response 80-Plex Human QuantiGene Plex Panel were normalized to the housekeeping control PPIB and data is displayed as log2 fold change over unstimulated control samples at the 3 different timepoints.
Click image to enlarge
Figure 7. Effect of LPS stimulation on gene expression in PBMCs.
Additionally, PBMC were stimulated with 5 µg/mL Phytohemagglutinin (PHA) and 5 µg/mL Concanavalin A (ConA) or 10 µg/mL LPS to measure the top ten induced and top ten repressed gene targets at 24h post simulation (see figure 8). Raw MFI data from the QuantiGene Plex assay using the Immune Response 80-Plex Human QuantiGene Plex Panel were normalized to the housekeeping control PPIB and data is displayed as log2 fold change over unstimulated control samples.
Click image to enlarge
Figure 8. Gene expression changes after 48h of stimulation with PHA, ConA or LPS.
NEW Invitrogen Immune Response 80-Plex Mouse QuantiGene Plex Panel
Analyze 80 cytokine, chemokine, and growth factor targets simultaneously to enable efficient immune response profiling, biomarker discovery, and validation.
Genes included in selected preconfigured 80-plex panels
Immune Response 80-Plex Human Plex Panel
CCL1 = I-309
CCL26 = Eotaxin-3
CXCL11 = I-TAC
HGF = HGF
IL18 = IL-18
IL3 = IL-3
LGALS3 = GAL-3
TNFSF10 = TRAIL
CCL13 = MCP-4
CCL4 = MIP-1b
CXCL13 = BLC
IFNA1 = INF-a
IL1A = IL-1A
IL31 = IL-31
LIF = LIF
TNFSF13 = APRIL
CCL17 = TARC
CCL7 = MCP-3
CXCL2 = MIP-2a
IFNG = INF-g
IL1B = IL-1B
IL34 = IL-34
LTA = TNF-b
TNFSF13B = BAFF
CCL19 = MIP-3b
CCL8 = MCP-2
CXCL5 = ENA-78
IL10 = IL-10
IL2 = IL-2
IL37 = IL-37
MIF = MIF
TREM1 = TREM-1
CCL2 = MCP-1
CD40LG = CD40L
CXCL6 = GCP-2
IL12A = IL-12p35
IL20 = IL-12p35
IL4 = IL-4
NGF = b-NGF
TSLP = TSLP
CCL21 = 6Ckine/SLC
CSF1 = M-CSF
CXCL9 = MIG
IL12B = IL12-p40
IL12B = IL12-p40
IL5 = IL-5
PTX3 = PTX3
VEGFA = VEGF-A
CCL22 = MDC
CSF2 = GM-CSF
CXCR3 = IP-10
IL13 = IL-13
IL13 = IL-13
IL6 = IL-6
TNF = TNF-a
PPIB
CCL23 = MPIF
CSF3 = G-CSF
FGF2 = FGF-2
IL15 = IL-15
IL15 = IL-15
IL8 = IL-8
TNFRSF12A = TWEAK
HPRT1
CCL24 = Eotaxin-2
CX3CL1 = Fractalkine
GZMA = Granzyme A
IL16 = IL-16
IL16 = IL-16
IL9 = IL-9
TNFRSF1B = TNF-R2
GAPDH
CCL25 = TECK
CCXL1 = GRO-a
GZMB = Granzyme B
IL17A = IL-17A
IL17A = IL-17A
KITLG = SCF
TNFRSF8 = CD30
GUSB
*QuantiGene panel (RNA) targets in red, QuantiGene housekeeping genes in blue, ProcartaPlex panel (protein) targets in black. Targets in ProcartaPlex panel that are NOT in QuantiGene panel are Eotaxin, IL-7, MIP-1a, MIP-3a, MMP-1.
Correlation data of Immune Response Human ProcartaPlex & QuantiGene Plex assays
PBMC were stimulated with 10 µg/mL Lipopolysaccharide (LPS) and correlation of RNA and protein expression was measured. Relative RNA and protein expression of ENA (CXCL5), GRO-alpha (CXCL1), MCP-3 (CCL7) and BLC (CXCL13) at 48h post stimulation with LPS is shown in figure 9. Raw MFI data from the Immune Response 80-Plex Human QuantiGene Plex were normalized to the housekeeping control PPIB. Protein data was acquired using the complementary Immune Response 80-Plex Human ProcartaPlex Panel. Data is displayed as normalized gene expression (RNA) and total amounts of protein (pg/mL) for unstimulated and LPS-stimulated samples at the 48h timepoint. RNA expression is represented by lines and protein expression by bars in the figure below.
Figure 9. Correlation of Gene (RNA) vs Protein expression at 3 different timepoints after stimulation with LPS.
To demonstrate the power of the 80-plex as a screening method, 30 samples were analyzed. 23 human serum samples were untreated, and 7 plasma samples were treated with LPS 100 ng/mL for 20 hours.
These were quantified in parallel using Invitrogen ProcartaPlex Human Immune Response Panel (80-plex). Differences in the expression pattern of 80 targets in untreated human serum samples and pre-treated plasma samples are shown in the following figures, grouped by low, medium, and high expression levels. Analytes are displayed on the x-axis, and mean concentration for untreated (blue lines) versus treated (orange lines) samples for each analyte on the y-axis.
Click image to enlarge
Click image to enlarge
Click image to enlarge
Figure 1. Differences in the expression pattern of 80 targets in untreated human serum samples and pre-treated plasma samples.
Additionally, a few analytes in the medium expressed samples are shown in more detail. These analytes showed the greatest degree of difference between treated versus untreated samples. This data is presented in four figures as demonstrated here.
Click image to enlarge
Figure 2. Concentration levels for a few analytes with medium expression levels.
To confirm scalability of ProcartaPlex multiplex immunoassays, human serum and stimulated plasma samples were run in parallel using both the Invitrogen ProcartaPlex Chemokine Panel 2 (9-plex) and the Invitrogen ProcartaPlex Human Immune Response Panel (80-plex). Experiments demonstrated high consistency between large-scale versus small-scale multiplex panels, verifying that high content screening can be performed without compromising the test results. Concentration levels for MCP-4 and CCL1 at different serum samples for small plex (9-plex) versus 80-plex is presented in the following figure. This was a method to demonstrate the level of correlation between small and big panels.
Click image to enlarge
Figure 3. Concentration levels for MCP-4 and CCL1 at different serum samples for small plex (9-plex) vs 80-plex.
To further demonstrate the degree of correlation between small and big plex, a regression analysis was performed. This yielded a R2 value >0.9. (0.99 for MCP-4, 0.93 for CCL1).
Click image to enlarge
Figure 4. Correlation Analysis between small and big plex.
Targets highlighted in bold are additional in the Immune Response 64-Plex Mouse ProcartaPlex Panel compared to the Immune Monitoring 48-Plex Mouse ProcartaPlex Panel (EPX480-20834-901).
The ability to scale the number of analytes and correlate data in multiplex experiments is critical for the progression of many projects. For example, it is not uncommon to begin a project by analyzing a large, comprehensive panel on few samples to determine which analytes are affected by a particular disease or drug treatment. The resulted list of analytes can then be investigated further with a smaller panel on a larger set of samples. Thus, there is a great need for scalability in multiplex panel assays to correlate data across different stages of investigation.
To demonstrate the scalability and reproducible performance regardless of plex size mouse serum and plasma samples of healthy, untreated mice were run in parallel on the ProcartaPlex Mouse Immune Monitoring Panel 48-Plex and the ProcartaPlex Mouse Immune Response Panel 64-Plex. Data in Figure 5 and 6 illustrates the high correlation for sample measurements for the same analytes in a large and small panel.
Click image to enlarge
Figure 5. Comparison data for selected analytes in blood samples from healthy, untreated mice.
Click image to enlarge
Click image to enlarge
Figure 6. Data illustrate the high correlation between large and small-scale ProcartaPlex panels.
Performance data from the largest High-Plex QuantiGene Plex assays
Immune Response 80-Plex Human QuantiGene Plex Panel
PBMC were stimulation with Lipopolysaccharide (LPS) and the gene expression change over time was measured with the Immune Response 80-Plex Human QuantiGene Plex. The majority of the 80 gene targets tested showed expression changes over unstimulated control samples with 10 µg/mL LPS-stimulation of PBMCs at 3 time points (see figure 7). Raw MFI data from the QuantiGene Plex assay using the Immune Response 80-Plex Human QuantiGene Plex Panel were normalized to the housekeeping control PPIB and data is displayed as log2 fold change over unstimulated control samples at the 3 different timepoints.
Click image to enlarge
Figure 7. Effect of LPS stimulation on gene expression in PBMCs.
Additionally, PBMC were stimulated with 5 µg/mL Phytohemagglutinin (PHA) and 5 µg/mL Concanavalin A (ConA) or 10 µg/mL LPS to measure the top ten induced and top ten repressed gene targets at 24h post simulation (see figure 8). Raw MFI data from the QuantiGene Plex assay using the Immune Response 80-Plex Human QuantiGene Plex Panel were normalized to the housekeeping control PPIB and data is displayed as log2 fold change over unstimulated control samples.
Click image to enlarge
Figure 8. Gene expression changes after 48h of stimulation with PHA, ConA or LPS.
NEW Invitrogen Immune Response 80-Plex Mouse QuantiGene Plex Panel
Analyze 80 cytokine, chemokine, and growth factor targets simultaneously to enable efficient immune response profiling, biomarker discovery, and validation.
Genes included in selected preconfigured 80-plex panels
Immune Response 80-Plex Human Plex Panel
CCL1 = I-309
CCL26 = Eotaxin-3
CXCL11 = I-TAC
HGF = HGF
IL18 = IL-18
IL3 = IL-3
LGALS3 = GAL-3
TNFSF10 = TRAIL
CCL13 = MCP-4
CCL4 = MIP-1b
CXCL13 = BLC
IFNA1 = INF-a
IL1A = IL-1A
IL31 = IL-31
LIF = LIF
TNFSF13 = APRIL
CCL17 = TARC
CCL7 = MCP-3
CXCL2 = MIP-2a
IFNG = INF-g
IL1B = IL-1B
IL34 = IL-34
LTA = TNF-b
TNFSF13B = BAFF
CCL19 = MIP-3b
CCL8 = MCP-2
CXCL5 = ENA-78
IL10 = IL-10
IL2 = IL-2
IL37 = IL-37
MIF = MIF
TREM1 = TREM-1
CCL2 = MCP-1
CD40LG = CD40L
CXCL6 = GCP-2
IL12A = IL-12p35
IL20 = IL-12p35
IL4 = IL-4
NGF = b-NGF
TSLP = TSLP
CCL21 = 6Ckine/SLC
CSF1 = M-CSF
CXCL9 = MIG
IL12B = IL12-p40
IL12B = IL12-p40
IL5 = IL-5
PTX3 = PTX3
VEGFA = VEGF-A
CCL22 = MDC
CSF2 = GM-CSF
CXCR3 = IP-10
IL13 = IL-13
IL13 = IL-13
IL6 = IL-6
TNF = TNF-a
PPIB
CCL23 = MPIF
CSF3 = G-CSF
FGF2 = FGF-2
IL15 = IL-15
IL15 = IL-15
IL8 = IL-8
TNFRSF12A = TWEAK
HPRT1
CCL24 = Eotaxin-2
CX3CL1 = Fractalkine
GZMA = Granzyme A
IL16 = IL-16
IL16 = IL-16
IL9 = IL-9
TNFRSF1B = TNF-R2
GAPDH
CCL25 = TECK
CCXL1 = GRO-a
GZMB = Granzyme B
IL17A = IL-17A
IL17A = IL-17A
KITLG = SCF
TNFRSF8 = CD30
GUSB
*QuantiGene panel (RNA) targets in red, QuantiGene housekeeping genes in blue, ProcartaPlex panel (protein) targets in black. Targets in ProcartaPlex panel that are NOT in QuantiGene panel are Eotaxin, IL-7, MIP-1a, MIP-3a, MMP-1.
Correlation data of Immune Response Human ProcartaPlex & QuantiGene Plex assays
PBMC were stimulated with 10 µg/mL Lipopolysaccharide (LPS) and correlation of RNA and protein expression was measured. Relative RNA and protein expression of ENA (CXCL5), GRO-alpha (CXCL1), MCP-3 (CCL7) and BLC (CXCL13) at 48h post stimulation with LPS is shown in figure 9. Raw MFI data from the Immune Response 80-Plex Human QuantiGene Plex were normalized to the housekeeping control PPIB. Protein data was acquired using the complementary Immune Response 80-Plex Human ProcartaPlex Panel. Data is displayed as normalized gene expression (RNA) and total amounts of protein (pg/mL) for unstimulated and LPS-stimulated samples at the 48h timepoint. RNA expression is represented by lines and protein expression by bars in the figure below.
Figure 9. Correlation of Gene (RNA) vs Protein expression at 3 different timepoints after stimulation with LPS.
Immune Response 80-Plex Human ProcartaPlex Panel
Immune Response 64-Plex Mouse ProcartaPlex Panel
