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Technical Information

Technical Notes: R&D Systems Is With You Every Step of the Way – Analyzing Proteins of Interest

Step 1 Use one of our Proteome Profiler™ Arrays as a screening tool to identify proteins of interest

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R&D Systems Human Cytokine Array Kit, Panel A (Catalog # ARY005) utilizes capture antibodies spotted onto a nitrocellulose membrane to allow high-throughput multi-analyte profiling of 36 cytokines, chemokines and acute phase proteins in a single sample. Chemiluminescent detection (left) and calculated pixel density histogram (right) show relative protein expression in human peripheral blood mononuclear cells incubated with PHA for 24 hours. Select proteins are highlighted for orientation. For a complete list of cytokines assayed in the Human Cytokine Array and the Mouse Cytokine Array, Panel A (Catalog # ARY006), please visit the Proteome Profiler page.

Step 2 Quantify your results with our dependable ELISA/Assay Kits

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GRO alpha in Peripheral Blood Mononuclear Cells (PBMC). PBMC were treated with PHA for 24 hours and assessed for CXCL1/GRO alpha using the Human CXCL1/GRO alpha Quantikine ELISA Kit (Catalog # DGR00). TNF-alpha and IFN-gamma in Activated T Cells. The production of TNF-alpha and IFN-gamma in the supernatant of PHA-activated human T cells (selected from peripheral blood mono­nuclear cells) are stimulated by CD3 activation using mouse anti-human CD3 monoclonal antibody (Catalog # MAB100). Cytokines were measured using human Quantikine ELISA kits for TNF-alpha (Catalog # DTA00C) and for IFN-gamma (Catalog # DIF50).

Step 3 Select from a wide range of antibodies and proteins to provide addi­tional information in many types of experimental procedures

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Intracellular Detection of TNF-alpha by Flow Cytometry. Human peripheral blood mononuclear cells treated with PMA and calcium ionomycin were stained with phycoerythrin (PE)-conjugated-mouse anti-human TNF-alpha monoclonal antibody (Catalog # IC210P; filled histogram). Staining with PE-conjugated isotype control (Catalog # IC002P; open histogram) highlights the specificity of the TNF-alpha antibody. TNF-alpha Immunocytochemistry in Human Peripheral Blood Mono­nuclear Cells (PBMC). TNF-alpha was detected in human PBMC using anti-human TNF-alpha/TNFSF1A polyclon­al anti­body (Catalog # AF-210-NA). Cells were stained using a Rhodamine Red™ X-conjugated anti-goat IgG secondary antibody (red) and counterstained with FluoroNissl™ green.

Rhodamine Red and FluoroNissl are trademarks of Molecular Probes, Inc.