RayBio® Human TNF alpha ELISA Kit for cell culture supernatants, plasma, serum, and lysate samples. 30 pg/ml sensitivity.
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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human |
| Sensitivity | 30 pg/ml Need more sensitivity? Check out the new NexaTag® kit for this target. Still not enough? Then your answer is our Ultrasensitive Biomarker Testing Service powered by Simoa™ technology. |
| Detection Range | 30 pg/ml - 6000 pg/ml |
| Specificity | This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human ANG, CD23, Eotaxin, GCSF, GM-CSF, GRO-alpha, GRO-beta, GRO-gamma, I-309, IFN-gamma, IL-1 alpha, IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p40), IL-12 (p70), IL-15, IL-16, IP-10, MCP-1, MCP-2, MCP-3, MCP-4, MCSF, MIG, MIP-1 alpha, MIP-1 beta, NAP-2, PDGF, PF-4. |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates |
| Recommended Dilution (Serum/Plasma) | 2 fold |
| Accession Number | P01375 |
| Gene Id | 7124 |
| Gene Symbols | TNF, TNFA, TNFSF2 |
| 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] |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Cardiovascular Disease, Cancer, Inflammation, Antibody Drug Development, Protein Therapeutics |
| Shipping Type | Blue ice |
| Storage | ≤-20°C |
Target Background
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.
Product Features
- 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
Standard Curves
Spiking & Recovery Results
| Sample Type | Average % Recovery | Range (%) |
|---|---|---|
| Serum | 94.53 | 82-103 |
| Plasma | 91.92 | 80-102 |
| Cell culture media | 96.43 | 84-104 |
Linearity Results
| Serum | Plasma | Cell Culture Media | ||
|---|---|---|---|---|
| 1:2 | Average % of expected Range (%) | 94 83-102 | 93 82-103 | 92 81-102 |
| 1:4 | Average % of expected Range (%) | 94 83-104 | 92 82-103 | 99 85-107 |
Typical Data
Kit Components
- 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
Additional 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
Assay Procedure Summary
- 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.
Storage/Stability
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.
