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Human IL-6 ELISA

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Citations (133)

The RayBiotech IL-6 ELISA kit is an in vitro enzyme-linked immunosorbent assay for the
quantitative measurement of human IL-6 in cell culture supernatants, plasma, serum, and lysate samples.

Lead time: Typically ships within 1-2 business days. No Friday shipments.

$420.00  
ELH-IL6-1
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Assay Format

Detection Range:
3 pg/ml - 1000 pg/ml
Recommended Dilution (serum & plasma):
2 fold
Sensitivity:
3 pg/ml

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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.
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

Protein Information

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)
Target Species:
  • Human

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

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

Standard Curves

Spiking & Recovery Results

Sample TypeAverage % RecoveryRange (%)
Serum92.6383-103
Plasma93.7484-104
Cell culture media95.6583-105

Linearity Results

  SerumPlasmaCell Culture Media
1:2Average % of expected
Range (%)		95
84-103		96
85-105		97
84-105
1:4Average % of expected
Range (%)		96
83-105		95
84-103		97
85-105

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
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Need your results faster? Try RayBiotech's SpeedELISA platform for quantitative detection in just three hours.

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.
  1. Thompson P., Shah A., Ntranos V., et al. Targeted Elimination of Senescent Beta Cells Prevents Type 1 Diabetes. Cell Metab . 2019 May 7;29(5):1045-1060.e10. doi: 10.1016/j.cmet.2019.01.021. Epub 2019 Feb 21.
    Species: Human
    Sample type: Conditioned Media (Islet conditioned media from islet cell cultures)
  2. Lu H., et al. A breast cancer stem cell niche supported by juxtacrine signalling from monocytes and macrophages. Nature Cell Biology (2014) doi:10.1038/ncb3041
    Species: Human
    Sample type: Conditioned Media (Mammary Epithelial Cell lines)
  3. Rathinam, Chozhavendan, et al. "Efficient differentiation and function of human macrophages in humanized CSF-1 mice." Blood 118.11 (2011): 3119-3128.
    Species: Human
    Sample type: Serum
  4. Rathinam, Chozhavendan, et al. "Efficient differentiation and function of human macrophages in humanized CSF-1 mice." Blood 118.11 (2011): 3119-3128.
    Species: Human
    Sample type: Conditioned Media (BM cells)
  5. Tian, Wenfang, et al. "Brahma?related gene 1 bridges epigenetic regulation of proinflammatory cytokine production to steatohepatitis in mice." Hepatology58.2 (2013): 576-588.
    Species: Human
    Sample type: Conditioned Media (HepG2 cells )
  6. Šenolt, Ladislav, et al. "Adipokine profile is modulated in subcutaneous adipose tissue by TNF? inhibitors in patients with rheumatoid arthritis." Annals of the rheumatic diseases 70.11 (2011): 2054-2056.
    Species: Human
    Sample type: Tissue Lysate (adipose tissue)
  7. Chai, Q., Lu, Z., Liu, Z. et al. Lung gene expression signatures suggest pathogenic links and molecular markers for pulmonary tuberculosis, adenocarcinoma and sarcoidosis. Commun Biol 3, 604 (2020). https://doi.org/10.1038/s42003-020-01318-0
    Species: Human
    Sample type: Conditioned media (Cell culture media)
  8. Said N., Sanchez-Carbayo M., Smith S., and Theodorescu D.RhoGDI2 suppresses lung metastasis in mice by reducing tumor versican expression and macrophage infiltration. J Clin Invest. Apr 2, 2012; 122(4): 1503-1518. doi: 10.1172/JCI61392
    Species: Human
    Sample type: Conditioned Media (Human UMUC3 coculture)
  9. Said N., Sanchez-Carbayo M., Smith S., and Theodorescu D.RhoGDI2 suppresses lung metastasis in mice by reducing tumor versican expression and macrophage infiltration. J Clin Invest. Apr 2, 2012; 122(4): 1503-1518. doi: 10.1172/JCI61392
    Species: Human
    Sample type: Tissue Lysate (Tumor lysate from lungs of baldder cancer)
  10. Said, Neveen, et al. "Tumor endothelin-1 enhances metastatic colonization of the lung in mouse xenograft models of bladder cancer." The Journal of clinical investigation 121.1 (2011): 132-147.
    Species: Human
    Sample type: Conditioned Media
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Great standard curve (R2=0.998). I used on supernatants and I suggest a 3 or 4 x dilution.
Anonymous
HMRI 03/03/2022, 11:37 AM
The package was delivered in perfect conditions. I tried the kit and my standard curve showed an R^2 =0.99. Unfortunately I didn't get the results from my samples because the signal was over saturated. I will repeat the experiment.
Gherardo
Houston Methodist Research Insitute 03/03/2022, 11:37 AM
We used Huh7.5 and A549 cells by infection with SARS-CoV-2 or transient ... activation by spike protein under our experimental conditions.
Dan
NanoBST 02/27/2022, 08:59 PM
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# Manual (Supernatants / Plasma / Serum)

# Manual (Lysates)

# SDS (Supernatants / Plasma / Serum)

# SDS (Lysates)

# Analysis Tool

Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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