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Pathway Focus: The JAK/STAT Pathway
Utilizing an alkyne-modified nucleoside, 5-ethynyl uridine (EU), and powerful click chemistry, newly synthesized RNA can be detected with a simple, two-step procedure. First, the alkyne-containing nucleoside is fed to cells or animals, and is actively and selectively incorporated into nascent RNA. Detection utilizes the chemoselective ligation or “click” reaction between an azide and an alkyne, where the modified RNA is detected with a corresponding azide-containing dye.
The JAK-STAT signaling pathway transmits information from chemical signals outside the cell to gene promoters in the nucleus, and affects cell proliferation, differentiation, and apoptosis.
Click-iT® RNA Alexa Fluor® Imaging Kits and HCS Assays provide the tools necessary to produce quantitative and qualitative measurements of RNA that arise from changes in DNA transcription.
The ability to detect newly synthesized RNA or changes in RNA levels will aid in understanding viral gene expression and genome replication, as well as its subcellular localization in host cells. The small size of the Click-iT® detection molecule allows easy penetration of complex samples. The assay is multiplex-compatible with other probes, including antibodies, for the simultaneous detection of RNA-interacting proteins.
|Description ||Cat. No. |
|Click-iT® RNA Alexa Fluor® 488 HCS Assay, 2-plate size ||C10327|
|Click-iT® RNA Alexa Fluor® 594 HCS Assay, 2-plate size ||C10328|
|Click-iT® RNA Alexa Fluor® 488 Imaging Kit, for 25 coverslips ||C10329 |
|Click-iT® RNA Alexa Fluor® 488 Imaging Kit, for 25 coverslips ||C10330|
In response to the 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.
IL-2, IL-3, IL-5, IL-7, IL-15, GM-CSF, thrombopoietin, and different growth hormones activate the STAT5 protein.
|Product||Cat. No.||Product||Cat. No.||Product||Cat. No. |
These 3-plex Luminex® assays provide a series of combined reagents for the simultaneous measurement of phosphorylated and total STAT1, STAT3, and STAT5a/b in cell lysates and tissue homogenates.
STAT proteins control transcription of specific genes in response to cytokine stimulation. Given that the diverse cellular response to IFN exposure is mediated by a common pathway, it is likely that a variety of STAT homo- and heterodimer combinations are responsible for each individual phenotype through differential gene regulation.
Differential activation of STAT1, STAT3, and STAT5a/b in different cell types suggests the possibility of different STAT homo- and heterodimer populations in response to interferon treatment. Significant activation of both STAT1 and STAT3 in TF-1 cells in response to IFN-α suggests a greater population of STAT1/STAT3 heterodimers. This is in contrast to TF-1 cell response to IFN-γ, where only STAT1 is significantly activated and STAT1 homodimers are the most likely transcriptional regulators.
This figure shows the advantages inherent to multiplex analysis for activation effects of cytokines. The ability to monitor activation of STAT1, STAT3, and STAT5a/b in a single well provides advantages in terms of time and sample requirements, and allows different markers in the same sample to be compared.
CellTrace™ Violet Cell Proliferation Assay—Violet Laser Cell Proliferation Reagents Make Analysis in GFP-Expressing Cells Possible
Like CFSE, the new Molecular Probes® CellTrace™ Violet Cell Proliferation Kit allows quantitation of cell generations by dye dilution using flow cytometry. Now with the violet excitation and blue emission of CellTrace™ Violet, you can perform proliferation experiments with a wide range of blue-excitable fluorophores, including GFP.
|Description ||Laser ||Ex/Em||Cat. No. |
|CellTrace™ Violet Cell Proliferation Kit ||405 nm ||405/450||V34557|
The JAK/STAT signaling pathway plays an important role in regulating cell proliferation, differentiation, and apoptosis. Upon binding of a variety of cytokines and growth factors to the appropriate receptors, JAK kinases are recruited and activated. JAKs then phosphorylate the receptors’ cytoplasmic domain, causing recruitment of STATs, which are in turn phosphorylated, dimerized, and translocated into the nucleus. Once in the nucleus, STAT family members (STAT1, 2, 3, 4, 5a, 5b, and 6) control transcription of specific genes in response to stimulation.
phosphoELISA™ kits allow you to measure the levels of total and phosphorylated JAK and STAT proteins with quantitative results in only half a day. phosphoELISATM kits provide a simple and unbiased way to quantify JAK/STAT protein levels with high specificity and better sensitivity than western blots. The kits come ready to use with all the necessary reagents, including recombinant standards that allow quantitative measurements. In addi¬tion, the flexible 96-strip well plate format allows you to run as many samples, or as few, as you need.
|Description ||Species ||Quantity||Cat. No. |
|STAT3 [pY705] ELISA Kit ||Hu, Ms, Rt ||96 tests||KHO0481|
|STAT5a [pY694] ELISA Kit ||Hu||96 tests||KHO0761|
|STAT5a (Total) ELISA Kit ||Hu||96 tests||KHO0751 |
|STAT5b [pY699] ELISA Kit ||Hu||96 tests||KHO5721|
|STAT6 [pY641] ELISA Kit ||Hu||96 tests||KHO0801 |
|JAK2 [pYpY1007/1008] ELISA Kit ||Hu||96 tests||KHO5621|
|JAK2 (Total) ELISA Kit ||Hu||96 tests||KHO5521|
We offer over 700 phosphorylation site–specific antibodies, and antibodies against extracellular markers to study the JAK/STAT and related pathways. This includes phosphorylation site–specific antibodies, which are tested on western blots with phosphorylation site–blocking peptides to ensure the phosphorylation specificity.
The JAK/STAT pathway is one of the crucial signaling pathways affecting cell functions, including proliferation, growth, hematopoiesis, and immune response. Our high-quality antibodies are ideal for experiments to detect total protein and even activated fractions. This information is valuable as scientist work toward understanding the role of each pathway protein.
Our JAK/STAT antibodies are validated not only in traditional techniques like western blotting and ELISA, but also in more challenging techniques like immunofluorescence, immunohistochemistry, and flow cytometry.
The availability of JAK/STAT antibodies produced with ABfinity™ recombinant rabbit monoclonal antibody technology enables researchers to have another layer of assurance in the quality and lot-to-lot consistency of these antibodies (STAT4 ABfinity™ Recombinant Rabbit Monoclonal Antibody and SUMO-3 ABfinity™ Recombinant Rabbit Monoclonal Antibody).