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Pathway Focus: The Tumor Necrosis Factor (TNF) Pathway
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Measure TNF-α Using Conventional ELISA—Invitrogen™ ELISA Kits and Antibody Pairs
Human tumor necrosis factor alpha (Hu TNF-α), also called cachectin, is a 157 amino acid cytokine mainly produced by monocytes and activated macrophages. In the central nervous system, TNF-α is involved in fever, anorexia, and alterations in pituitary hormone release. In the cardiovascular system, TNF-α plays a role in shock, acute respiratory distress, and capillary leakage syndrome (procoagulation). TNF-α is instrumental in the process of acute tubular necrosis and nephritis in the kidney and ischemia, colitis, and hepatic necrosis in the gastrointestinal system. Major TNF-α effects on metabolism include LPL suppression, protein catabolism, lipid catabolism, ACTH release, and insulin resistance. | TNF-α is defined by its participation in inflammatory activities, cell toxicity, increased NK function, and mediation of TNF-α tumor toxicity. The ability to precisely measure TNF-α would provide important information in many of these research areas. Invitrogen™ ELISA kits offer an easy and reliable way to quantify the amount of TNF-α in your serum, plasma, or supernatant sample. Precoated ELISA kits are available for a variety of species including human, mouse, rat, swine, and monkey, and economy pack sizes are available for your convenience. For more savings, matched Antibody Pairs are also available.
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| Figure 1. Invitrogen™ ELISA kits offer an easy, reliable method for quantifying TNF-α. | | ) | | Figure 2. Lot-to-lot consistency of the TNF-α ELISA Kit. Individual production lots were analyzed using 4 levels of control specimens (colors), ensuring low variations between lots (<20%). |
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| Product | Species | Range | Sample Vol. Needed | Incubation Time | Quantity | Cat. No. |
|---|---|---|---|---|---|---|
| ELISA Kits | | | | |||
| TNF-α Human ELISA Kit | Human | 15.6–1,000 pg/mL | 100 µL | 4 hr | 96 tests 192 tests 480 tests | KHC3011 KHC3012 KHC3011C |
| TNF-α Ultrasensitive Human ELISA Kit | Human | 0.5–32 pg/mL | 50 µL | 3 hr | 96 tests 192 tests 480 tests | KHC3014 KHC3013 KHC3014C |
| TNF-α Mouse ELISA Kit | Mouse | 19.5–1,250 pg/mL | 50 µL | 2.5 hr | 96 tests 192 tests 480 tests | KMC3011 KMC3012 KMC3011C |
| TNF-α Rat ELISA Kit | Rat | 15.6–1,000 pg/mL | 50 µL | 2.75 hr | 96 tests 192 tests 480 tests | KRC3011 KRC3012 KRC3011C |
| TNF-α Monkey ELISA Kit | Monkey | 15.6–1,000 pg/mL | 50 µL | 4 hr | 96 tests 192 tests | KPC3011 KPC3012 |
| TNF-α Swine ELISA Kit | Swine | 23–1,500 pg/mL | 25 µL | 4 hr | 96 tests 192 tests | KSC3011 KSC3012 |
| Antibody Pairs (user flexibility) | ||||||
| TNF-α Human Antibody Pair | Human | NA | NA | NA | 10 plates | CHC1753 |
| TNF-α Mouse Antibody Pair | Mouse | NA | NA | NA | 10 plates | CMC3013 |
| TNF-α Rat Antibody Pair | Rat | NA | NA | NA | 10 plates | CRC3013 |
| TNF-α Swine Antibody Pair | Swine | NA | NA | NA | 10 plates | CSC1753 |
Profiling TNF Pathway Biomarkers—Invitrogen™ Multiplex Assays for Luminex® xMAP® Technology
The TNF superfamily of cytokines includes a multifunctional group of proinflammatory cytokines that activate signaling pathways for cell survival, apoptosis, inflammatory responses, and cellular differentiation. | In response to inflammation and infection, the proinflammatory cytokine TNF-α is produced by macrophages, lymphocytes, fibroblasts, and keratinocytes. Invitrogen™ multiplex bead immunoassays were developed for simultaneous detection of activated proteins to provide a more complete picture. |
| Figure 3. Ultra-low detection of cytokines. Comparison of detection levels between the conventional Invitrogen™ Human Cytokine 10-Plex Panel and the Invitrogen™ Human Cytokine Ultrasensitive 10-Plex Panel. |
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| Description | Premixed Panel | Species | Cat. No. | |
| Apoptosis 3-Plex Panel | Cytochrome c, Caspase-3 [175/176], PARP [214/215] | Human | LHO0007 |  |
| Cytokine Human 10-Plex Panel | GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-α + Buffer kit | Human | LHC0001 | |
| Cytokine Human 25-Plex Panel | Eotaxin, GM-CSF, IFN-α, IFN-γ, IL-1β, IL-1RA, IL-2, IL-2R, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40/p70), IL-13, IL-15, IL-17, IP-10, MCP-1, MIG, MIP-1α, MIP-1β, RANTES, TNF-α + Buffer kit | Human | LHC0009 | |
| Cytokine Human 30-Plex Panel | EGF, Eotaxin, FGF-basic, G-CSF, GM-CSF, HGF, IFN-α, IFN-γ, IL-1β, IL-1RA, IL-2, IL-2R, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40/p70) IL-13, IL-15, IL-17, IP-10, MCP-1, MIG, MIP-1α, MIP-1β, RANTES, TNF-α, VEGF + Buffer kit | Human | LHC6003 | |
| Cytokine Mouse 10-Plex Panel | GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p40/p70), TNF-α + Buffer kit | Mouse | LMC0001 | |
| Cytokine Mouse 20-Plex Panel | FGF basic, GM-CSF, IFN-γ, IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IL-12 (p40/p70), IL-17, IP-10, KC, MCP-1, MIG, MIP-1α, TNF-α, VEGF + Buffer kit | Mouse | LMC0006 | |
| Cytokine Ultrasensitive Human 10-Plex Panel | GM-CSF, IFN-γ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNF-α + Buffer kit | Human | LHC6004 | |
| Death Receptor Human 3-Plex Panel | TNF-RI, TNF-RII, DR5 + Buffer kit | Human | LHC0006 | |
| Inflammatory Cytokine Human 5-Plex Panel | GM-CSF, IL-1β, IL-6, IL-8, TNF-α + Buffer kit | Human | LHC0003 | |
| Inflammatory Cytokine Mouse 4-Plex Panel | GM-CSF, IL-1β, IL-6, TNF-α + Buffer kit | Mouse | LMC0003 | |
| Th1/Th2 Cytokine Monkey 5-Plex Panel | IFN-γ, IL-2, IL-4, IL-10, TNF-α + Buffer kit | Monkey | LPC0001 | |
| TNF-RI Human Singleplex Bead Kit | Human | LHC3021 | | |
| TNF-RII Human Singleplex Bead Kit | Human | LHC3031 | | |
| TNF-α Human Singleplex Bead Kit | Human | LHC3011 | | |
| TNF-α Monkey Singleplex Bead Kit | Monkey | LPC3011 | | |
| TNF-α Mouse Singleplex Bead Kit | Mouse | LMC3011 | | |
| TNF-α Rat Singleplex Bead Kit | Rat | LRC3011 | | |
| TNF-α Ultrasensitive Human Singleplex Bead Kit | Human | LHC3013 | |
TNF-α–Induced Degradation of IκBα—Recombinant TNF-α for Cell Culture Applications
TNF-α is a proinflammatory cytokine that activates various signaling pathways, including the NF-κB signaling pathway. NF-κB, a regulator of the expression of genes for both innate and adaptive immune reponses, is localized in the cytoplasm bound to IκB. | Upon TNF-α stimulation, IκBα is phosphorylated, which marks the protein for degradation, making NF-κB free to translocate to the nucleus to induce gene expression. We offer bioactive recombinant TNF-α for use in cell culture applications. Mouse TNF-α, rat TNF-α, and swine TNF-α are also available in multiple pack sizes. |
| Figure 4. Jurkat cells treated with recombinant human TNF-α. The level of phosphorylated IκBα increased upon TNF-α stimulation. As expected, the total level of IκBα decreased upon stimulation due to proteosomal degradation. IκBα [pS32] and IκBα (total) levels were measured using the IκBα [pS32] Human ELISA Kit and IκBα (Total) Human ELISA Kit, respectively. |
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| Description | Quantity | Cat. No. | |
| Recombinant Human TNF-α | 5 µg | PHC3015 | |
| Recombinant Human TNF-α | 50 µg | PHC3016 | |
| Recombinant Human TNF-α | 100 µg | PHC3011 | |
| Recombinant Human TNF-α | 1 mg | PHC3013 | |
Definitive Tools for Investigating Autophagy—Premo™ Autophagy Sensors
The role autophagy plays in TNF-induced cell death is not yet completely defined. Autophagy, a cell death process involved in cancer, neurodegeneration, and immunological disorders, has been challenging to study because of a lack of accurate, sensitive, and robust assays. New Premo™ Autophagy Sensors allow researchers to view this process in live cells for the first time. Based on LC3B, a critical component and indicator of autophagy, Premo™ Autophagy Sensors combine the selectivity of an LC3B–fluorescent protein (FP) fusion with the transduction efficiency of second-generation BacMam technology, enabling unambiguous visualization of this protein. | The BacMam system enables efficient transduction in a wide variety of cells, including neurons and neural stem cells, using an easy, one-step protocol. To visualize autophagy, simply add BacMam LC3B-FP to the cells and incubate overnight for protein expression. Using these kits, researchers can now detect autophagy temporally and spatially using fluorescence microscopy and high-content imaging. |
| Figure 5. Detecting autophagy with Premo™ Autophagy Sensor LC3-RFP. U2OS cells were transduced with Premo™ Autophagy Sensor LC3-RFP and Cellular Lights™ MAP4-GFP. The following day, cells were treated with 50 µM chloroquine. Twenty-four hours later, cells were incubated with Hoechst 33342 before imaging. |
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| Description | Quantity | Cat. No. | |
| Premo™ Autophagy Sensor LC3B-GFP | 1 kit | P36235 | |
| Premo™ Autophagy Sensor LC3B-RFP | 1 kit | P36236 | |
For research use only. Not intended for human or animal therapeutic or diagnostic use.
