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Product Description
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit, 10 Plate Kit |
| Estimated Lead Time | 1-2 business days |
| Species | Human |
| Sensitivity | 3 pg/ml |
| Detection Range | 3 pg/ml - 1000 pg/ml |
| Specificity | This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF. |
| Compatible Sample Types | Cell Culture Supernatants, Plasma, Serum, Tissue Lysates, Cell Lysates |
| Recommended Dilution (Serum/Plasma) | 2 fold |
| Accession Number | P05231 |
| Gene Id | 3569 |
| Gene Symbols | IL6 |
| Protein Name / Synonyms | Interleukin-6 (IL-6) (B-cell stimulatory factor 2) (BSF-2) (CTL differentiation factor) (CDF) (Hybridoma growth factor) (Interferon beta-2) (IFN-beta-2) |
| Quantitative/Semi-Quantitative | Quantitative |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Angiogenesis, Cardiovascular Disease, Cancer, Inflammation |
| Shipping Type | Blue ice |
| Storage | ≤-20°C |
Introduction
The IL-6 ELISA employs an antibody specific for human IL-6 coated on a 96-well plate. Standards and samples are pipetted into the wells and IL-6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human IL- 6 antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IL-6 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Interleukin 6 (IL-6) is a pleiotropic cytokine that regulates cell growth and differentiation of various tissues, and plays an important role in the immune response and acute phase reactions. IL-6 acts as both a pro-inflammatory and anti-inflammatory cytokine. It is secreted by T cells and macrophages to stimulate an immune response, e.g., during infection and after trauma, especially burns or other tissue damage leading to inflammation. IL-6's role as an anti-inflammatory cytokine is mediated through its inhibitory effects on TNF-alpha and IL-1, and activation of IL-1R alpha and IL-10.
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 | 92.63 | 83-103 |
| Plasma | 93.74 | 84-104 |
| Cell culture media | 95.65 | 83-105 |
Linearity Results
| Serum | Plasma | Cell Culture Media | ||
|---|---|---|---|---|
| 1:2 | Average % of expected Range (%) | 95 84-103 | 96 85-105 | 97 84-105 |
| 1:4 | Average % of expected Range (%) | 96 83-105 | 95 84-103 | 97 85-105 |
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.
