IMMUNOLOGY Exploring Signaling LUNARIS™ Chemokine Kits

Human 11-Plex: CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CXCL1, CXCL8 (IL-8), CXCL10 (IP-10), CXCL12 (SDF1A)

Mouse 12-Plex: CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CCL22, CXCL1, CXCL10 (IP-10), CXCL12 (SDF1A), CX3CL1 (Fractalkine)

Chemokines

Chemokine receptor

Plasma membrane its | 2019.05 | 08694 | www.vierviertel.com | 08694 | 2019.05 its Cytoplasm

K Chemokine AYOXXA Intracellular signaling

Chemotaxis

www.ayoxxa.com LUNARIS™ Human & Mouse Chemokine Kits Validated, scalable, robust

Validated for quantitative analysis of up to 12 different in one sample

Profile homeostatic and proinflammatory chemokine signatures in sample volumes as low as 3 µL

Scalable and standardized assay architecture with robust chemistry guarantees reproducibility

Harmonized panels for human and murine samples allow rapid translation of results from lab to clinic

LUNARIS™ Human 11-Plex Chemokine Kit Quantifies CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CXCL1, CXCL8 (IL-8), CXCL10 (IP-10), CXCL12 (SDF1A) in serum and cell culture supernatant.

LUNARIS™ Mouse 12-Plex Chemokine Kit Quantifies CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CCL22, CXCL1, LUNARIS™ Human 11-Plex CXCL10 (IP-10), CXCL12 (SDF1A,) CX3CL1 (Fractalkine) Chemokine Kit in serum and cell culture supernatant. For Research Use Only. Not for use in diagnostic procedures.

Chemokine Signaling

Chemokines constitute the largest family of and mediate versatile and powerful signaling in cell migration, Analytes contained in LUNARIS™ Chemokine Kits homeostasis, , , embryogenesis, grouped by cellular function cancer metastasis and several other medically relevant functions.

The four classes of chemokines initiate complex signaling cascades by binding to specific 7-transmembrane G - Mononuclear Granulocyte coupled receptors. traffic cell traffic traffic

Building fine-tuned concentration gradients, chemokines exert precise control of diverse regulatory pathways and cause pleiotropic effects that extend far beyond their originally described role as leucocyte chemoattractants. A plethora of literature bears witness to the known importance of these small cytokines in a vast number of biological functions, while a growing number of publications hints at a yet undescribed complex interplay between target cells and their respective Th1, Th2, Th17 cells , , chemokines. natural killer cells

Chemokine repertoires of some signaling axes are extensively conserved between mice and humans, making murine models the system of choice in more than just immunology contexts. CCL20, CCL22, CCL2, CCL3, CCL4, CCL11, CXCL1, CX3CL1, CXCL10 CCL5, CCL19, CX3CL1 CXCL8 (IL-8) Relevant research areas:

Inflammation and infection biology Note: CXCL12 is mainly involved in homing and autoimmune disease (e.g., retention of granulocytes, B cells & precursors). Hematopoiesis and angiogenesis

www.ayoxxa.com 2 Excellent Data Quality Validation of LUNARIS™ Human 11-Plex Chemokine Kit*

Table 1: Performance Analyte LoD LLOQ ULOQ DR Figure 1: Performance Median assay parameters CCL2 0.2 0.32 200 3.7 Bar diagram depicting the CCL2 CCL3 from 20 experiments. CCL3 4.6 12 300 3.0 median assay parameters CCL4 L0D = Limit of detection, CCL4 2.9 4.8 600 2.6 shown in table 1. The orange CCL5 CCL11 CCL5 1.4 8 1000 3.5 LLOQ = Lower limit of bars represent the dynamic CCL19

quantification, CCL11 2.3 7,2 900 3.5 range of the assay from LoD Analytes CCL20 CCL19 10.7 24 15000 4.2 CXCl1 ULOQ = Upper limit (left border) to the ULOQ CXCL8 of quantification, CCL20 5.1 24 600 3.3 (right border). The LLOQ is CXCL10 CXCL1 9.2 40 1000 3.5 CXCL12 DR = Logarithmic detection depicted as a white line. 1 1 0 10 CXCL8 0.3 0.48 300 2.3 0. 10 1000 0000 range. All values reported Measurements from 20 1 CXCL10 4.9 8 3000 3.5 concentration [pg/mL ] in pg/mL. CXCL12 24.2 120 3000 4.2 validation experiments.

Figure 2: Accuracy Figure 3: Precision Median recovery rate 200 Between-run precision (BRP) 40 from 10 experiments of QC samples spiked into cell quantifying markers in 150 culture supernatants. Each data 30 %) %)

quality control samples y( point represents the median

100 P( 20 spiked at two different ver between-run precision of BR

concentrations into cell eco two different QC sample culture supernatants. R 50 concentrations measured in 10 The targeted recovery 10 experiments. The targeted range (70–130%) is 0 precision (<20%) is denoted 0 3 5 0 2 2 3 4 5 0 2 2 4 11 19 L1 L8 11 19 20 L1 L8 20 L1 L1 CL CL CL CL C C L1 L1 CL CL CL C C by the dotted lines/grey CL CL CL C C denoted by the dotted CL C C C CL CL CL X X C C C C C C X X C C C C C C X X C C C C C X X lines/grey shaded area. C C shaded area. C C

* Validation data of the corresponding mouse kit are available upon request.

Innovative technology for Multiple options to access translational proteomics the advantages of LUNARIS™

Ease of readout and handling Complete testing and readout services Fully integrated system Send in samples for testing and receive a complete analysis report Flexible scalability from low to high throughput Perform assays in your lab, then send in Readout of 384 samples in less than one hour completed assay plates for readout

AYOXXA’s proprietary LUNARIS™ platform for multiplex protein Innovative panel development analysis is a fully integrated and scalable system. A dedicated Custom panel configuration reader automates image-based analysis of immunoassay beads in a planar array, so every bead is read and every bead Custom multiplex assay development counts.

LUNARIS™ is optimized for sample volumes as low as 3 µL – one-tenth the volume required for similar technologies – yet allows full multiplex testing without compromising data quality or precision from precious samples.

With advantages of superior data quality, workflow flexibility and conservation of precious samples, LUNARIS™ enables reliable quantification of biomarkers from model to man – from lab to clinic – from data to insight.

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Ordering information * Chemokine Kit Human 11-Plex LUNARIS™ Kits Chemokine Kit Mouse 12-Plex Measured induplicate www.ayoxxa.com [email protected] +49 Call theexperts: signaling chemokine Exploring technology for translational proteomics innovative multiplex protein AYOXXA’s www.ayoxxa.com ( 0 ) 221-222 529-0 4 ×96 1 ×96 BioChips of No. 4 ×96 1 ×96 LHCK-20110F LHCK-20110S Cat. No. Cat. LMCK-20120F LMCK-20120S samples of No. 184 40 184 40

* from datafrom to insight. modelfrom to man lab –from to clinic biomarkers of quantification reliable enables quality, and efficiency, flexibility LUNARIS™ studies. advantages its With in terms of translating knowledge generated in clinical protein analysis platform is optimized for applications. Our multiplex LUNARIS™ panelsassay for translational research platform, our mission is to develop robust Building technology innovative an upon research. vision of enabling success in translational Biosystems AYOXXA E-Mail: [email protected] E-Mail: 529-0 (0) 222 +49 Phone: 221 50829 1 Nattermannallee Cologne BioCampus GmbH Biosystems AYOXXA HEADQUARTERS GERMANY Köln, Germany is dedicated to the E-Mail: [email protected] E-Mail: Phone: 02142 MA Cambridge, 1 Broadway Inc. Biosystems AYOXXA USA +1 (617) 5545 599 – – USA

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