BioPath Online

Pathway Focus: The Toll-Like Receptor Pathway

Antibodies for the Toll-Like Receptor Pathway
Antibodies for the Toll-Like Receptor Pathway—Novel Recombinant Antibodies for the Highest Consistency
Recombinant Proteins for Toll-Like Receptor Signaling ResearchRecombinant Proteins for Toll-Like Receptor Signaling Research—Recombinant Human IL-33

Ultrasensitive Cytokine and Chemokine DetectionUltrasensitive Cytokine and Chemokine Detection—Reliably Measure Low-Expression Inflammatory Markers After Toll-Like Receptor Activation
Uncover Subtle Changes in Cytokine LevelsUncover Subtle Changes in Cytokine Levels—Ultrasensitive Multiplex Cytokine Assays for Luminex® xMAP® Technology
New Antibodies

New Immunoassays

New Molecular Probes® Products

Antibodies for the Toll-Like Receptor Pathway—Novel Recombinant Antibodies for the Highest Consistency

  • Measure total and phosphorylated proteins
  • Unmatched lot-to-lot consistency with new ABfinity™ technology
  • High sensitivity and specificity

Toll-like receptor (TLR) signaling plays an essential role in the innate immune response. Activation of TLR signaling through recognition of pathogen-associated molecular patterns leads to the transcriptional activation of genes encoding proinflammatory cytokines, chemokines, and costimulatory molecules, which subsequently control the activation of the antigen-specific adaptive immune response.
We offer one of the most extensive antibody portfolios to study the TLR pathway, including phosphorylation site–specific antibodies for detection of specific phosphorylation status. Invitrogen antibodies are produced under ISO conditions with the highest quality controls. Our new ABfinity™ technology is used to generate the highest lot-to-lot consistency by cloning the antibody genes and expressing them in mammalian cells. The ABfinity™ platform is used to generate many of the antibodies in our TLR portfolio.

  • Learn More about Phosphospecific Antibodies

Flow cytometric detection of Jurkat cells
(click to enlarge)

Figure 1. Flow cytometric detection of Jurkat cells labeled with JNK1-2 [pTpY183/185] ABfinity™ antibody. Jurkat cells were stimulated with 25 μg/mL anisomycin, then fixed and permeabilized using FIX & PERM® reagents. Cells were then stained with the JNK1-2 antibody in the absence (blue trace) or presence (red trace) of the phosphopeptide immunogen, followed by detection using Alexa Fluor® 488 goat anti–rabbit IgG. The black trace represents unstimulated cells.

Immunohistochemistry analysis
(click to enlarge)

Figure 2. Immunohistochemistry analysis of human squamous lung, breast, and gastric carcinoma and mesothelioma tissues labeled with JNK1-2 [pTpY183/185] ABfinity™ antibody. FFPE human squamous lung (top left), breast (top right), gastric carcinoma (bottom left), and mesothelioma (bottom right) tissues were labeled with JNK1-2 [pTpY183/185] ABfinity™ Recombinant Rabbit Monoclonal Antibody (2 µg/mL). Tissues were pretreated with EDTA and detected with the SuperPicture™ Polymer DAB Kit. Images were taken at 40x magnification. Note nuclear staining in tumor cells.

Western blot of HEK lysates 
Figure 3. Western blot of HEK lysates labeled with JNK1-2 [pTpY183/185] ABfinity™ antibody. JNK1-2 [pTpY183/185] ABfinity™ Recombinant Rabbit Monoclonal Antibody (0.1 µg/mL) was used to label JNK1/2 in untreated HEK lysates (lane 1) or anisomycin-treated HEK lysates (lane 2).


DescriptionReactivity
Applications
Quantity
 Cat. No.
 
IRAK4 ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Sw, Sh, Rt, Qu, Eq, Cn, B)
F, IF/ICC
100 µg
700026
IR/IGF1R [pY1162/pY1163] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (X, Rt, Ms, Mk, Cp, Ch, Cn, B, Eq)
E, WB
100 µg700393
AKT [pS473] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Ms, Hu (Z, X, Rt, Mk, Ma, Eq, Fe, Eq, Cp, Ch, Cn, B)
E, WB,F, IHC, IF/ICC
100 µg700392
ERK1/2 [pT185/pY187] ABfinity™ Recombinant Rabbit Monoclonal Antibody
Hu (Z, X, Rt, Ms, Cp, Ch, B)
E, WB, IHC
100 µg700012
JNK1-2 [pTpY183/185] ABfinity™ Recombinant Rabbit Monoclonal Antibody (clone D12H7L17)
Hu (Z, X, Sw, Rt, Or, Ne, Ms, Mk, Gf, Eq, Cp, Ch, Cn, B)
E, WB,F, IHC, IF/ICC
100 µg700031
MEK3/6 [pSpT189/193]/[207/211], Rabbit Polyclonal Antibody - UNCONJ
Hu, Ms
WB
100 µg44470
MEK4 [pSpT257/261], Rabbit Polyclonal Antibody -UNCONJ
Hu (Ms)
WB100 µg44474G
TLR4, Rabbit Polyclonal Antibody - UNCONJ
Hu
WB, E
100 µg363700
TLR5 (C-term), Rabbit Polyclonal Antibody - UNCONJ
Hu
WB, E
100 µg363900

Reactivity: B = bovine; Ch = chicken; Cn = canine; Cp = chimpanzee; Eq = equine; Fe = feline; Gf = goldfish; Ha = hamster; Hu = human; Ne = nematode; Rb = rabbit; Rt = rat; Ms = mouse; Sh = sheep; Sw = swine; Ma = mammalian; Mk = monkey (rhesus); Or = orangutan; P = primate; Qu = quail; X = Xenopus; Z = zebrafish. ( ) indicates reactivity predicted but not tested.

Recombinant Proteins for Toll-Like Receptor Signaling Research—Recombinant Human IL-33

  • High biological activity—more results with less protein
  • High purity—no interference from other proteins or contaminants
  • Freeze/thaw stability—reproducible data from one experiment to the next
  • Proven compatibility—validated for use in cell culture using GIBCO® media

In addition to our extensive antibody portfolio for detecting Toll-like receptor (TLR) signaling proteins, we now offer the human IL-33 recombinant protein for TLR signaling studies.
It has been demonstrated that interleukin-33 (IL-33) affects the response of macrophages to TLR ligands such as LPS, and plays a role in the regulation of TLR signaling. IL-33, a member of the IL-1 family of cytokines, binds to the ST2 receptor, a member of the Toll-like/IL-1 receptor superfamily.

Members of this superfamily share a common intracellular domain, the Toll/interleukin-1 receptor (TIR) domain.


Description
Qty.
Cat. No.

Recombinant Human IL-33
10 µg
PHC9254
Recombinant Human IL-33
25 µgPHC9255
Recombinant Human IL-33100 µgPHC9251
Recombinant Human IL-33
1 mgPHC9253

Ultrasensitive Cytokine and Chemokine Detection—Reliably Measure Low-Expression Inflammatory Markers After Toll-Like Receptor Activation

  • Quantitate low-expressed IL-1β, IL-6, IL-8, and TNF-α
  • Ultrasensitive ELISA kits are 20x more sensitive than conventional ELISAs
  • Simple assay procedure

Microbes that penetrate an epithelial barrier and enter a tissue site are encountered by the three types of sentinel immune cells: tissue macrophages, mast cells, and immature dendritic cells. These cells must be able to distinguish between apoptotic particles generated by normal tissue turnover and particles that are indicative of infection. The molecules mostly responsible for making the distinction are part of the Toll-like receptor (TLR) family. Stimulation of macrophages or mast cells through their TLRs leads to the secretion of proinflammatory cytokines (such as IL-1 and TNF-α) and lipid mediators, thereby initiating the inflammatory response.  

Now you can reliably quantitate the inflammatory markers IL-1β, IL-6, IL-8, and TNF-α at levels not detectable by conventional ELISAs. Our Ultrasensitive ELISA Kits allow for the ability to detect low-expressed proteins and are 10x more sensitive than conventional ELISAs. The assay procedure is no different from any other sandwich ELISA kit, so it’s simple to perform.
Greater sensitivity of ultrasensitive ELISA compared to conventional ELISA
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Figure 1. Greater sensitivity of Ultrasensitive ELISA compared to conventional ELISA. Representative standard curves show that the Ultrasensitive IL-1β ELISA Kit is about 10x more sensitive than the conventional ELISA.



DescriptionSpecies
Quantity Cat. No.

IL-1β Ultrasensitive ELISA Kit
Human
96 tests
KHC0014
Order 
IL-6 Ultrasensitive ELISA Kit
Human96 tests
KHC0064Order
IL-8 Ultrasensitive ELISA Kit
Human96 tests
KHC0084Order
TNF-α Ultrasensitive ELISA Kit
Human96 tests
KHC3014Order

Uncover Subtle Changes in Cytokine Levels—Ultrasensitive Multiplex Cytokine Assays for Luminex® xMAP® Technology

  • Ultrasensitive detection—uncover subtle changes that could be missed
  • Small sample volume—conserve precious samples
  • Speed—saves time and labor
  • Minimal variation—get reliable results, lot after lot

Cytokines comprise a diverse group of small proteins that mediate signals between cells. Cytokines play important roles in normal developmental processes such as B and T cell maturation, vasculogenesis, and organogenesis. Cytokines also act systemically, providing signals from sites of injury or inflammation to potentially distant responding tissues.

Under certain circumstances, such as response to bacterial infection, cytokines can be detected in biological fluids at relatively high concentrations. In chronic disease states, however, disease-associated cytokines may be present in relatively lower concentrations in biological fluids. 
Numerous reports provide evidence that the accurate quantification of relatively low levels of cytokines can be achieved by ultrasensitive immunoassays. These reports also provide evidence that the detection of subtle changes in cytokine concentrations can provide valuable information about disease states.

The Ultrasensitive multiplex Luminex® kits for GM-CSF, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IFN-γ, and TNF-α allow sensitive quantitative measurements of these soluble proteins in serum, plasma, and tissue culture supernatants. Ultrasensitive assays detect cytokines in the femtogram range, allowing you to uncover subtle changes. Furthermore, Ultrasensitive assays require only small sample volumes and thus conserve your precious samples. And because we have over 20 years of assay development expertise, you can be confident that Invitrogen™ multiplex Luminex® assays accurately measure your target of interest, lot after lot.


Ultralow detection of the 10 human cytokines
(click to enlarge)
 
Figure 1. Ultralow detection of 10 human cytokines. Least detectable dose values seen using the Invitrogen™ Human Cytokine 10-plex and the Invitrogen™ Human Cytokine 10-plex Ultrasensitive Luminex® panel.


DescriptionQuantity
Cat. No.

Human Cytokine 10-plex Ultrasensitive Kit
(GM-CSF, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IFN-γ, TNF-α + buffer kit)
100 tests
LCH6004Order