25 pg/ml - 20000 pg/ml
Recommended Dilution (serum & plasma):
This ELISA kit shows no cross-reactivity with any of the cytokines tested: rat CINC-2, CINC-3, CNTF, Fractalkine, IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-10, GM-CSF, IFN-gamma, Leptin, Lix, MCP-1, MIP-3 alpha, beta- NGF, TIMP-1, VEGF.
Number of Targets Detected:
Compatible Sample Types:
- Cell Culture Supernatants
- Cell Lysates
- Tissue Lysates
Method of Detection:
1, 2, or 5 x 96-Well Strip Microplate Kit
Protein Name & Synonyms:
Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Tumor necrosis factor alpha (TNF alpha), also known as lymphotoxin and cachectin, is a pleiotropic cytokine. TNF alpha is a smaller cytokine with a molecular weight of 26 kDa (transmembrane) and 17 kDa (soluble) that binds to the same receptors producing a vast array of effects similar to those of TNF-beta. Both TNF-alpha and TNF-beta share 30% amino acid homology and have similar biological activities. TNF-alpha is produced primarily by monocytes, neutrophils and macrophages. It is found in synovial cells and macrophages in the tissues. TNF-alpha and TNF-beta can modulate many immune and inflammatory functions, while having the ability to inhibit tumor growth. Target tumor cells must express TNF receptors 1 (55 kDa) and 2 (75 kDa) to be killed, with the p55 receptor mediating the cytotoxic response.
- Strip plates and additional reagents allow for use in multiple experiments
- Quantitative protein detection
- Establishes normal range
- The best products for confirmation of antibody array data
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Stop Solution
- Assay Diluent(s)
- Lyophilized Standard
- Biotinylated Detection Antibody
- Streptavidin-Conjugated HRP
- TMB One-Step Substrate
Other Materials Required
- Distilled or deionized water
- Precision pipettes to deliver 2 µl to 1 µl volumes
- Adjustable 1-25 µl pipettes for reagent preparation
- 100 µl and 1 liter graduated cylinders
- Tubes to prepare standard and sample dilutions
- Absorbent paper
- Microplate reader capable of measuring absorbance at 450nm
- Log-log graph paper or computer and software for ELISA data analysis
- Prepare all reagents, samples and standards as instructed in the manual.
- Add 100 µl of standard or sample to each well.
- Incubate 2.5 h at RT or O/N at 4°C.
- Add 100 µl of prepared biotin antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared Streptavidin solution to each well.
- Incubate 45 min at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
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The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.