Enhance your research with Invitrogen's cell biology publications—BioProbes® , ProbesOnline™, and BioPath Online™.
Pathway Focus: TheToll-Like Receptor (TLR) Pathway
Gibco® cytokines and growth factors are used in a wide variety of applications including cell stimulation, proliferation, and differentiation. Gibco® cytokines and growth factors have been bioassayed in cells cultured using Gibco® media. So if you’re already using Gibco® media, you can have more confidence in the compatibility of these reagents in your cell culture–based assays.
Toll-like receptor signaling plays an essential role in the innate immune response. Pathogen-associated molecular patterns are recognized by toll-like receptors. The signaling cascade initiated by binding leads to the transcriptional activation of genes encoding specific pro-inflammatory cytokines, which control the activation of an antigen-specific immune response.
Activation of toll-like receptor signaling through IRF7 induces IFNα and IFNβ, promoting an antiviral immune response. Activation of toll-like receptor signaling through AP-1 and NF-κB induces IL-1β, IL-6, IL-8, IL-12, MIP-1α, MIP-1β, RANTES, and TNFα, mediating an inflammatory response.
Life Technologies produces a selection of Gibco® cytokines and growth factors to aid in immunology research. To help ensure Gibco® growth factors are of the highest quality, each protein is analyzed for purity along with structural homogeneity to ensure a biologically active protein is produced
The LIVE/DEAD® Viability/Cytotoxicity Kit is a quick and easy, two-color assay to determine viability of cells in a population. The kit contains two probes: calcein AM and ethidium homodimer-1. Calcein AM is a fluorogenic esterase substrate that is hydrolyzed to a green-fluorescent product (calcein); thus green fluorescence is an indicator of cells that have esterase activity as well as an intact membrane to retain the esterase products.
Ethidium homodimer-1 is a high-affinity, red-fluorescent nucleic acid stain that is only able to pass through compromised membranes of dead cells.
The LIVE/DEAD® Viability/Cytotoxicity kit can detect cytotoxicity following activation of toll-like receptors. By clear discrimination between live and dead cells in a population, researchers will be able to quickly and easily assess cytotoxicity following activation of toll-like receptors.
|Product ||Cat. No. |
|LIVE/DEAD® Viability/Cytotoxicity Kit ||L3224|
These 5-plex kits provide a series of combined reagents for the simultaneous measurement of human IL-1β, IL-6, IL-8, TNF-α, and GM-CSF in serum, plasma, or tissue culture supernatant. The assay may be run alone or in combination with other singleplex assay kits from Invitrogen.
Note: Human Inflammatory Cytokine 5-Plex Panel reagents are intended for use in the Luminex® 100™ or 200™ System only.
Human Inflammatory Cytokine Magnetic 5-Plex Panel reagents are intended for use in the Luminex® 100™⁄ 200™ or FlexMAP® 3D Systems only.
Assay validation includes bead and protein specificity, precision (intra- and inter-assay, 10%), parallelism between recombinant and natural samples, linearity of dilution, recovery (>80%), and comparison to ELISA (>90%). Premixed Multiplex Kits include General Buffer Reagent Kit components and Antibody Bead Kit components, with reagents premixed for the number of markers in the assay.
Toll-like receptor (TLR) signaling plays an essential role in the innate immune response. TLR signaling leads to the transcriptional activation of genes encoding for pro-inflammatory cytokines, chemokines, and co-stimulatory molecules, which subsequently control the adaptive immune response.
Invitrogen™ multiplex assays utilize Luminex® technology to enable reliable quantification of multiple human cytokines on the TLR/cytokine pathway (IL-1β, IL-6, IL-8, and TNF-α). Singleplex assays for macrophage inflammatory proteins (MIP-1α and MIP-1β) are also available. These assays can be effectively conducted in serum, plasma, or tissue culture supernatant.
Multiplex analysis kits enable the simultaneous detection and quantification of multiple molecules activated through TLR signaling. The ability to monitor these key molecules in a single well provides user advantages in terms of time, sample type requirements, and data collection and analysis.
The Alexa Fluor® 488–Annexin V Apoptosis Detection Kits detect apoptotic cells using recombinant annexin V conjugated to the Alexa Fluor® 488 dye, and dead cells using propidium iodide (PI). After treatment with both probes, apoptotic cells fluoresce green, dead cells fluoresce red and green, and live cells show little or no fluorescence.
There is a growing body of evidence that links toll-like receptor signaling with pro-apoptotic activities, perhaps as part of innate and adaptive immune responses in various organisms. The targeted Alexa Fluor® 488 dye conjugate identifies apoptotic cells by binding to phosphatidyl serine (PS) that has been translocated from the inner leaflet to the outer leaflet of the plasma membrane (a hallmark of apoptosis). Because the Alexa Fluor® 488 dye is brighter and more photostable than fluorescein (FITC), it provides better separation and population resolution. Propidium iodide is included in the kit as a dead-cell stain.
These kits allow researchers to make rapid flow cytometric–based assessments of apoptotic versus dead cells as part of their toll-like receptor pathway research.
|Description ||Size||Cat. No. |
|Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 and PI, 50 assays for flow cytometry ||1 kit||V13241|
|Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 and PI, 250 assays for flow cytometry ||1 kit||V13245|
|Annexin Binding Buffer, 5X concentrate for flow cytometry||1 kit||V13246|