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Human IL-6 ELISA by RayBiotech Inc.

Manufacturer RayBiotech Inc.  |   Model: ELH-IL6
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RayBio® Human IL-6 ELISA Kit for cell culture supernatants, plasma, and serum samples.


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Human IL-6 ELISA
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Antigen Information

Accession Number: P05231

Gene ID: 3569

Gene Symbols: IFNB2; 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

Assay Format

Detection Range: 3 pg/ml - 1000 pg/ml

Recommended Dilution (serum & plasma): 2 fold

Sensitivity: 3 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.

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 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

 

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.

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.