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Enhance your research with Invitrogen's cell biology publications—BioProbes® , ProbesOnline™, and BioPath Online™.


Immunology Catalog
Cell and Tissue Analysis

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.




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
SpeciesRangeSample Vol. Needed
 Incubation TimeQuantityCat. 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.