Human TNF-alpha ELISA by RayBiotech Inc.
Place your order directly with the manufacturer.
Antigen Information
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]
Target Species: Human
Assay Format
Detection Range: 30 pg/ml - 6000 pg/ml
Recommended Dilution (serum & plasma): 2 fold
Sensitivity: 30 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
Number of Targets Detected: 1
Species Detected: Human
Compatible Sample Types: Cell Culture Supernatants; Cell Lysates; Plasma; Serum; Tissue Lysates
Design Principle: Sandwich-based
Method of Detection: Colorimetric
Quantitative/Semi-Quantitative: Quantitative
Solid Support: 96-well Microplate
Product Specifications
Size: 1, 2, or 5 x 96-Well Strip Microplate Kit
Product Features
Additional Notes
- 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
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 |
Application Notes
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
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
Protocol Outline
- 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.
Product Overview
