BioPath Online

Share |

Gibco® Cytokines and Growth Factors Involved in the Toll-Like Receptor Pathway

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



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

For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Gibco® Cytokine and Growth Factor Selection Guide

Product Cat. No.
Product
Cat. No. Product
Cat. No.
IFNα
PHC4814IL-6
PHC0064
MIP-1β
PHC1034Z
IFNβ
PHC4244IL-8
PHC0884
RANTES
PHC1054
IL-1β
PHC0815MIP-1α   
PHC1104TNFα
PHC3015

Viability Detection Following Toll-Like Receptor Activation

  • Simple to perform—add → incubate → analyze; no wash steps required
  • Versatile—compatible with adherent cells, nonadherent cells, and certain tissues; use fluorescence microscopy, flow cytometry, HCS or fluorimetry, or a fluorescence microplate
  • Simple to analyze—green-fluorescent cells are live; red-fluorescnt cells are dead



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.

Live and dead kangaroo rat (PtK2) cells stained with ethidium homodimer-1 and the esterase substrate calcein AM
Live and dead kangaroo rat (PtK2) cells stained with ethidium homodimer-1 and the esterase substrate calcein AM.

Both are provided in our LIVE/DEAD® Viability/Cytotoxicity Kit. Live cells fluoresce a bright green, whereas dead cells with compromised membranes fluoresce red-orange.
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Ordering information

Product
Cat. No.
LIVE/DEAD® Viability/Cytotoxicity Kit
L3224

Detect and Quantify TLR Pathway Markers—Luminex® Multiplex Assays

  • Superior results—accurate, reproducible, and sensitive quantitation of multiple TLR pathway markers
  • Fast and easy—multiple tests per sample enable data collection and analysis typically in less than one day
  • High quality—in-house manufactured antibodies help ensure consistent high quality and excellent specificity and sensitivity



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.

  • Learn More about Luminex® Multiplex Technology
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Ordering Information

Description
Reactivity
Markers
Cat. No.
Human Inflammatory Cytokine Magnetic 5-Plex Panel
Human
IL-1β, IL-6, IL-8, TNF-α, and GM-CSF
LHC0003M
Human Inflammatory Cytokine 5-Plex Panel
Human
IL-1β, IL-6, IL-8, TNF-α, and GM-CSF
LHC0003

Detect Apoptosis Associated With Toll-Like Receptor Pathway Activation

  • Better data—the use of Alexa Fluor® 488 dye produces brighter, more photostable Annexin V conjugates
  • Simple conversion—Alexa Fluor® 488 dye easily fits into the same filter configuration as FITC
  • Convenience—ready-to-use assays with validated flow cytometry protocols
  • More economical—reduced price per test for larger and smaller kit formats



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.

Apoptosis detection by flow cytometry

Apoptosis detection by flow cytometry. 
Camptothecin-induced Jurkat cells were stained with the recommended concentration of annexin V conjugate for 30 min, then washed, resuspended, and counterstained with propidium iodide. Cells were analyzed on the Attune™ Acoustic Focusing Cytometer using a 488 nm laser source and 530/30 nm and 575/24 nm emission filters.
Ordering Information

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
For research use only. Not intended for any animal or human therapeutic or diagnostic use. © 2010 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners.  Luminex, FlexMAP, 100, and 200 are trademarks of Luminex Corporation.