ELISpot MAX™ Deluxe Set Human IL-6

Pricing & Availability
Regulatory Status
RUO
Other Names
Interleukin 6, B-cell Stimulatory Factor 2 (BSF-2), CTL Differentiation Factor (CDF), Hybridoma Growth Factor
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Product Citations
publications
A. ELISpot-MAX_Human-IL-6_1_1128223
Human peripheral blood mononuclear cells (PBMC) at 2.5 x 103 cells/well were incubated for 20 hours in the absence or presence of 100 ng/mL LPS. The number of cells secreting Human IL-6 was analyzed by ELISpot.
  • A. ELISpot-MAX_Human-IL-6_1_1128223
    Human peripheral blood mononuclear cells (PBMC) at 2.5 x 103 cells/well were incubated for 20 hours in the absence or presence of 100 ng/mL LPS. The number of cells secreting Human IL-6 was analyzed by ELISpot.
  • B.
ELISpot-MAX_Human-IL-6_2_1128223
    Human PBMC (1 x 104 cells/mL) were incubated for 20 hours in the absence or presence of either 1X Cell Activator without Brefeldin A (Cat. No. 423301) or 1X Cell Activator with Brefeldin A (Cat. No. 423303). BioLegend’s Cell Activator cocktail contains PMA and ionomycin at the optimized concentration. Brefeldin A functions to inhibit intracellular protein transport. As a result, it inhibits secretion of cytokines such as IL-6. The number of cells secreting Human IL-6 was analyzed by ELISpot.
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430524 5 plates $615
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Description

Human IL-6 (Interleukin), also known as B-cell Stimulatory Factor 2 (BSF2), Interferon beta-2 (IFN-β-2), and Hybridoma Growth Factor, is a cytokine which is expressed by many different cell types such as macrophages, T-cells, osteoblasts, endothelial cells, and hepatocytes. It is involved in both pro-inflammatory and anti-inflammatory responses. In particular, IL-6 has critical roles in multiple different biological processes including, but not limited to B-cell maturation, osteoclast formation, nerve cell differentiation, muscle healing and hypertrophy. Thus, abnormal levels of expression and activity of IL-6 can result in a wide range of diseases from chronic inflammation, cancer, to neurodegenerative diseases.

BioLegend’s ELISpot MAX™ Deluxe Set contains pre-optimized essential reagents and additional buffers and solution for Enzyme-Linked Immunosorbent Spot assay. This kit is designed for detection of human IL-6 secreting cells. A monoclonal antibody specific for human IL-6 is coated on ethanol-activated 96-well PVDF membrane plate as the capture. Stimulated and/or unstimulated cells are pipetted directly into the wells, and then incubated at 37°C for a specific period of time. During this period, the capture antibody binds to human IL-6 from the secreting cells in the vicinity. Then, the cells are removed and a biotinylated anti-human IL-6 detection antibody is added, which is then followed by Avidin-HRP. The TMB membrane substrate is subsequently added. The TMB membrane substrate reacts with HRP to form a blue-black precipitate at the sites of human IL-6 secreting cells. The end result is visible spots, which correspond to footprints of IL-6 secreting cells, on the membrane surface.

Product Details
Technical data sheet

Kit Contents

Kit Contents
  • Human IL-6 ELISpot MAX™ Capture Antibody (200X) 
  • Human IL-6 ELISpot MAX™ Detection Antibody (200X) 
  • Human IL-6 Standard (used as positive control) 
  • Avidin-HRP (1000X) 
  • TMB Membrane Substrate
  • Coating Buffer A (5X) 
  • Assay Diluent B (5X) 

Product Details

Verified Reactivity
Human
Application

ELISpot

Materials Not Included
  • PVDF-bottom plate: Millipore MultiScreenHTS IP Filter Plate (Cat. No. MSIPS4W10) is recommended. 
  • Wash Buffer: BioLegend Cat. No. 421601 is recommended, or PBS + 0.05% Tween-20  
  • Plate Sealers: BioLegend Cat. No. 423601 is recommended 
  • 0.22 μm filters
  • Sterile PBS (Phosphate-Buffered Saline): 8.0 g NaCl, 1.16 g Na2HPO4, 0.2 g KH2PO4, 0.2 g KCl, add deionized water to 1 L; pH to 7.4, 0.22 μm filtered
  • Sterile cell culture media
  • 37°C CO2 incubator
  • Deionized (DI) water 
  • Adjustable pipettes to measure volumes ranging from 2 μL to 1 mL 
  • Wash bottle, manifold dispenser, or automated microplate washer 
  • Tubes for dilution
  • Timer 
  • Stereomicroscope or an automated ELISpot reader

Antigen Details

Cell Type
Endothelial cells, Macrophages, Osteoblasts, T cells
Biology Area
Cancer Biomarkers, Immuno-Oncology, Immunology, Innate Immunity, Neurodegeneration, Signal Transduction
Molecular Family
Cytokines/Chemokines, Innate Immune Signaling
Gene ID
P05231 View all products for this Gene ID
UniProt
View information about IL-6 on UniProt.org

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Go To Top Version: 1    Revision Date: 11/28/2023

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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