BioPath Online

Pathway Focus: The Tumor Necrosis Factor (TNF) Pathway

Measure TNF-α Using Conventional ELISA
Measure TNF-α Using Conventional ELISA—Invitrogen™ ELISA Kits and Antibody Pairs
Profiling Tumor Necrosis Factor (TNF) Pathway BiomarkersProfiling TNF Pathway Biomarkers—Invitrogen™ Multiplex Assays for Luminex® xMAP® Technology

TNF-α–Induced Degradation of IκBαTNF-α–Induced Degradation of IκBα—Recombinant TNF-α for Cell Culture Applications
Definitive Tools for Investigating AutophagyDefinitive Tools for Investigating Autophagy—Premo™ Autophagy Sensors
 

New Antibodies

New Immunoassays

New Molecular Probes® Products

Measure TNF-α Using Conventional ELISA—Invitrogen™ ELISA Kits and Antibody Pairs

  • Easy to use—simple, reliable assays
  • Versatile—quantify TNF-α in serum, plasma, or supernatant
  • Compatible—kits available for a variety of species

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.

  • View all ELISA and Antibody Pair Products



Figure 1. Invitrogen™ ELISA kits offer an easy, reliable method for quantifying TNF-α.

 
Lot-to-lot consistency of the TNF-α ELISA Kit

 
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%).


Product
Species RangeSample Vol. Needed
 Incubation TimeQuantity 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 PairMouse
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

  • Superior performance—accurate, reproducible, and sensitive quantitation of TNF pathway proteins
  • High quality—antibodies manufactured in-house ensure excellent specificity and sensitivity
  • Fast and easy protocols—perform and analyze your data in less than one day

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.


DescriptionPremixed Panel
SpeciesCat. No.

Apoptosis 3-Plex Panel
Cytochrome c, Caspase-3 [175/176], PARP [214/215]
Human
LHO0007
Order 
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
LHC0001Order
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
LHC0009Order
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
LHC6003Order
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
LMC0001Order
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
LMC0006Order
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
LHC6004Order
Death Receptor Human 3-Plex Panel
TNF-RI, TNF-RII, DR5 + Buffer kit
Human
LHC0006Order
Inflammatory Cytokine Human 5-Plex Panel
GM-CSF, IL-1β, IL-6, IL-8, TNF-α + Buffer kit
Human
LHC0003Order
Inflammatory Cytokine Mouse 4-Plex Panel
GM-CSF, IL-1β, IL-6, TNF-α + Buffer kit
Mouse
LMC0003Order
Th1/Th2 Cytokine Monkey 5-Plex Panel
IFN-γ, IL-2, IL-4, IL-10, TNF-α + Buffer kit
Monkey
LPC0001Order
TNF-RI Human Singleplex Bead Kit

Human
LHC3021Order
TNF-RII Human Singleplex Bead Kit

Human
LHC3031Order
TNF-α Human Singleplex Bead Kit

Human
LHC3011Order
TNF-α Monkey Singleplex Bead Kit

Monkey
LPC3011Order
TNF-α Mouse Singleplex Bead Kit

Mouse
LMC3011Order
TNF-α Rat Singleplex Bead Kit

Rat
LRC3011Order
TNF-α Ultrasensitive Human Singleplex Bead Kit

Human
LHC3013Order

TNF-α–Induced Degradation of IκBα—Recombinant TNF-α for Cell Culture Applications

  • 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 with our media systems—one less variable for you to worry about

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.


DescriptionQuantity
 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

  • Versatile—detect autophagy in live or formaldehyde-fixed cells
  • Ready to use—easy, one-step protocol (add to cells and incubate overnight)
  • Content-rich data—combine with other live-cell probes (Organelle Lights™, Cellular Lights™, MitoTracker®, and LysoTracker® reagents) or antibodies

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.


DescriptionQuantity
Cat. No.

Premo™ Autophagy Sensor LC3B-GFP
1 kit
P36235Order 
Premo™ Autophagy Sensor LC3B-RFP
1 kit
P36236Order

For research use only.  Not intended for human or animal therapeutic or diagnostic use.