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Test Autoimmune Responses With ProtoArray® Microarrays and ProtoPlex™ Assays

  • Profile over 9,000 targets in a day
  • Custom testing for any auto-antibody targets
  • Minimal serum needed for each assay

The ProtoArray® Human Protein Microarray is the first high-density microarray and contains thousands of unique, full length human proteins including kinases, phophatases, GPCRs, nuclear receptors, and proteases, spotted in duplicate on a thin, 1 in x 3 in nitrocellulose-coated glass slide. ProtoArray® Human Protein Microarray version 5.0 contains over 9,000 unique human proteins individually purified and arrayed under native conditions to maximize functionality. The arrays are an ideal tool for comparing two sample groups to find novel autoimmune responses .

The ProtoPlex™ Immune Response Assay service is a multiplex, plate-based assay that couples proteins to Luminex® microspheres and can be used to characterize up to 50 autoantibody responses. Choose antigens from our catalog of more than 9,000 expression-ready human clones or supply your own content. The high throughput of the ProtoPlex™ assay allows researchers to gather qualitative immune response data from hundreds of samples in a single experiment.

Together, these platforms will help researchers identify new autoimmune responses using minimal sample sizes, and then quickly verify if those markers should be pursued for downstream development.

 

Class V5 Content
 Protein Kinases (unique) 268
 Protein Kinases (splice variants, mutants, etc.) 776
 Transcription factors 328
 Membrane proteins 2635
 Nuclear proteins 2252
 Signal transduction 1526
 Secreted proteins 192
 Cell communication 1687
 Metabolism 3862
 Cell death 505

 Table 1. ProtoArray® microarrays feature over 9,000 human full-length human proteins.

For Research Use Only. Not for use in diagnostic procedures.

TGF-β Helps Moderate Autoimmunity

  • High biological activity
  • High purity
  • Proven compatibility with Gibco® media


In a recent publication, Surh and Sprent (Nat Immunol 13:628-30 (2012)) present results of an investigation using knockout mice to examine the role of TGF-β in autoimmunity. Their work demonstrates that TGF-β is likely to be essential to prevent homeostatic proliferation by self antigens from developing into autoimmunity. Life Technologies offers high-quality Gibco® TGF-β for use in autoimmunity research.

Gibco® Cytokines and Growth Factors are high-purity recombinant proteins with high bioactivity. Gibco® growth factors are of the highest quality because each protein is analyzed for purity along with structural homogeneity to help ensure a biologically active protein is produced. And Gibco® Cytokines and Growth Factors have been bioassayed with Gibco® media. Get your cytokines and growth factors from the media company you use and trust.


Ordering Information

Product Cat. No.  Cat. No.
 10 µg100 µg
TGF-β1 Recombinant HumanPHG9214PHG9211
TGF-β2, Recombinant HumanPHG9124PHG9121
   
 5 µg250 µg
TGF-β3 Recombinant HumanPHG9305PHG9302

For Research Use Only. Not for use in diagnostic procedures.

Measure Cytokines That Regulate Autoimmunity

  • Quantitative, reliable protein measurement
  • Consistent results
  • Easy-to-run assays—4 hours of incubation time


Complex interactions of various cytokines play key roles in the regulation of autoimmune responses. Rheumatoid arthritis, multiple sclerosis, and insulin-dependent diabetes are three major human autoimmune disorders considered to be typical T cell–mediated diseases. There are a number of cytokines that regulate and control autoimmune responses. For example, IL-23 and IL-12 production (in the absence of any infection) may be important players in autoimmune inflammation. Proinflammatory cytokines such as IL-6 and IL-17, as well as IL-4 and TGFB may ameliorate autoimmunity, depending on the timing and level of production.

The ELISA or multiplexing bead based assays using the Luminex® platform are reliable and easy methods to obtain quantitative measurements of cytokine proteins. The assay procedure and quantitative results can be quickly achieved within a day in a 96 well plate format.

 

 Figure 1. The range of the Human IL-23 ELISA kit is 62.5 to 4,000 pg/mL. The recombinant protein standard in the assay kit mimics native proteins (supernatant of THP-1 cells induced with LPS).

For Research Use Only. Not for use in diagnostic procedures.

Endocytosis Detection and Role of TLR3 in Autoimmunity

  • Optimized probes for detection of endocytosis in live cells
  • 10,000 MW dextran conjugates available in red- and green-fluorescent forms
  • Useful for live cell fluorescence-imaging, high-content screening (HCS), flow cytometery, high-throughput screening (HTS) on platforms such as the Tali™, FLoid®, and Attune® Systems


Our proprietary pH-sensitive Molecular Probes® pHrodo™ conjugates of dextran, E. coli, S. aureus, and Zymosan particles are designed to be specific sensors of endocytosis. We’ve developed both green- and red-fluorescent forms to fit with the excitation sources and filters in most instruments.

The dye is nonfluorescent at neutral pH and fluoresces bright red or green acidic environments. And because pHrodo™ dye is minimally fluorescent at neutral pH, the need for wash steps and quencher dyes in the assay is dramatically reduced.

pH-sensitive pHrodo™ dextrans and other pHrodo™ conjugates are useful for the study of endocytosis and to investigate autoimmunity other pathways. For example, Skold and colleagues used pHrodo™ dye–conjugated dextrans to investigate the relationship between endocytosis and TLR3 activation (Blood 120:768–777 (2012)). 

 

 

Figure 2. Detection of endocytosis in A431 cells using pHrodo™ Red dextran, 10,000 MW. A431 macrophage cells were plated in complete medium and cultured overnight. The following day, cells were rinsed once with Live Cell Imaging Solution. pHrodo™ Red dextran, 10,000 MW (left) or pHrodo™ Green dextran, 10,000 MW (right) was added to cells at 100 µg/mL, and plates moved back to 37°C for 30 min. NucBlue™ Live cell stain was used as DNA counterstain. 

For Research Use Only. Not for use in diagnostic procedures.

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