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Pathway Focus: The Toll-Like Receptor Pathway
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Antibodies for the Toll-Like Receptor Pathway—Novel Recombinant Antibodies for the Highest Consistency
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.
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) (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. |
) (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. |
 | | 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). |
| Description | Reactivity | 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 µg | 700393 | |
| 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 µg | 700392 | |
| ERK1/2 [pT185/pY187] ABfinity™ Recombinant Rabbit Monoclonal Antibody | Hu (Z, X, Rt, Ms, Cp, Ch, B) | E, WB, IHC | 100 µg | 700012 | |
| 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 µg | 700031 | |
| MEK3/6 [pSpT189/193]/[207/211], Rabbit Polyclonal Antibody - UNCONJ | Hu, Ms | WB | 100 µg | 44470 | |
| MEK4 [pSpT257/261], Rabbit Polyclonal Antibody -UNCONJ | Hu (Ms) | WB | 100 µg | 44474G | |
| TLR4, Rabbit Polyclonal Antibody - UNCONJ | Hu | WB, E | 100 µg | 363700 | |
| TLR5 (C-term), Rabbit Polyclonal Antibody - UNCONJ | Hu | WB, E | 100 µg | 363900 | |
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
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 µg | PHC9255 | |
| Recombinant Human IL-33 | 100 µg | PHC9251 | |
| Recombinant Human IL-33 | 1 mg | PHC9253 | |
Ultrasensitive Cytokine and Chemokine Detection—Reliably Measure Low-Expression Inflammatory Markers After Toll-Like Receptor Activation
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. |
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| Description | Species | Quantity | Cat. No. | |
| IL-1β Ultrasensitive ELISA Kit | Human | 96 tests | KHC0014 | |
| IL-6 Ultrasensitive ELISA Kit | Human | 96 tests | KHC0064 | |
| IL-8 Ultrasensitive ELISA Kit | Human | 96 tests | KHC0084 | |
| TNF-α Ultrasensitive ELISA Kit | Human | 96 tests | KHC3014 | |
Uncover Subtle Changes in Cytokine Levels—Ultrasensitive Multiplex Cytokine Assays for Luminex® xMAP® Technology
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. |
) (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. |
| Description | Quantity | 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 | LCH6004 | |
