Pathway Focus: The JAK/STAT Pathway
The Tali™ Image Cytometer combined with the Tali™ Apoptosis Kit offers the following benefits:
- Accuracy–obtain statistically significant analysis comparable to flow cytometry
- Visualization–the Tali™ instrument and kit generate quantitative, image-based analysis of cell populations undergoing apoptosis
- Convenience–the kit contains all reagents necessary for the fluorescent detection of apoptotic and necrotic cells
The Tali™ Image Cytometer is a 3-channel (i.e., bright field, green fluorescence, and red fluorescence) benchtop assay platform that uses state-of-the-art optics and sophisticated digital image analysis to perform assays on cells in suspension, including assays for apoptosis, GFP and RFP expression, cell viability (live, dead, and total cells), and cell counts.
The JAK/STAT pathway is theorized to play a major role in hematopoietic cancers (i.e., leukemia, lymphoma, and myeloma) and thus has been a target for clinical research efforts. Current studies have focused on developing inhibitors of several key pathway elements to promote apoptosis. Using Jurkat cells (lymphoblasts from an acute T cell leukemia patient) as a model system, we have developed a robust and quantitative apoptosis assay for the Tali™ Image-Based Cytometer.
|The Tali™ Image Cytometer offers two-color fluorescence quantitative analysis for routine endpoint assays, such as two-color apoptosis assays, which are unavailable with a quick microscope check. In addition, it is the ideal companion instrument for flow cytometry workflows, allowing confirmation of critical parameters before setting up more complicated flow cytometry experiments.
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Tali™ Image Cytometer Ordering Information
- ABfinity™ rabbit recombinant antibodies for sensitive detection of STAT6
- Enables consistent performance for your assays
- Consistent product from lot to lot
ABfinity™ recombinant rabbit antibodies are produced from specific recombinant clones to enable consistent antibody performance over time. ABfinity™ antibodies are available against many targets, including JAK/STAT pathway proteins.
STAT6 (signal transducers and activators of transcription 6) belongs to the STAT family of proteins which, in response to cytokines and growth factors, are phosphorylated by receptor-associated kinases and act as transcription factors. When induced by CD80 or CD86 ligand binding, phosphorylated CD28 recruits and activates STAT6, which in turn translocates to the nucleus and binds to a STAT6-responsive element on the human IL-4 promoter. STAT6 is crucial for IL-4–mediated biological responses, and it is also involved in processes mediated by IL-13, a cytokine related to IL-4. IL-4 and IL-13 have been shown to induce the production of IgE, a major mediator in an allergic response.
We offer an extensive antibody portfolio to study the JAK/STAT signaling pathway:
- Learn more about ABfinity™ Recombinant Rabbit Monoclonal Antibodies
- Use the Primary Antibody Search Tool
|Figure 1. Immunocytochemistry analysis of HeLa cells detected with STAT6 ABfinity™ Recombinant Rabbit Monoclonal Antibody and using (A) Alexa Fluor® 488 goat anti-rabbit as the secondary antibody (green), (B) DAPI as the nuclear stain (blue), and (C) Alexa Fluor® 594 phalloidin to stain actin (red). (D) A composite image of cells shows cytoplasmic localization of STAT6.|
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|STAT6 ABfinity™ Recombinant Rabbit Monoclonal Antibody – Purified||100µg||Order Now|
Gibco® recombinant proteins offer:
- High biological activity
- High purity
- Proven compatibility with Gibco® media
STAT proteins are a family of cytoplasmic signaling proteins involved in signal transduction of several cytokines and growth factors. In response to a cytokine or growth factor binding to cell surface receptors, STAT proteins become activated by tyrosine phosphorylation. Phosphorylated STAT proteins subsequently move to the cell nucleus, where they activate transcription.
Individual cytokines are thought to activate specific STAT family members. For example, STAT1 and STAT2 play a central role in the interferon signaling cascade. The binding of interleukin-6 (IL-6) family cytokines and some other growth factors triggers STAT3 phosphorylation. IL-2, IL-3, IL-5, IL-7, IL-15, GM-CSF, thrombopoietin, and different growth hormones activate the STAT5 protein.
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.
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- Easy-to-run ELISA–4 hours of incubation time
- Convenient precoated, removable 8-well strips
- Quantitative, sensitive results
STAT5 protein is part of the JAK/STAT family of proteins. Phosphorylated STAT5 is typically present in healthy breast epithelia. Loss of STAT5 phosphorylation is frequent in cancer progression.
JAK/STAT regulates cell proliferation, differentiation, and apoptosis. Various cytokines may initiate JAK activation, which then causes downstream phosphorylation of STAT proteins that ultimately translocate into the nucleus to control transcription of genes. Identification of inhibitors of the JAK/STAT protein family is a key focus in cancer research.
The STAT5 ELISA Kits provide a great screening tool for drug inhibitors by enabling easy, reliable measurement of phosphorylated STAT5 protein in cell lysates. The assay can be carried out within a day in a 96-well plate format, with quantitative results. Convenient strip-well plates also provide flexibility to run as few samples as needed without wasting material.
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|Figure 2. High specificity of the phosphoELISA™ Kit—no cross-reactivity. HEL cell lysates were incubated with the capture antibody used in the STAT5a [pY694] ELISA (lane 2). An antibody specific for STAT5a and STAT5b was used as a positive control (lanes 1 and 4). IgG beads were used as a negative control (lane 3). The capture antibody recognizes the a isoform of STAT5 but not the b isoform. Thus, the STAT5a [pY694] ELISA does not cross-react with the STAT5b protein.|