The HTRF human GM-CSF kit is designed for the quantification of human GM-CSF release in cell supernatant.

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  • No-wash No-wash
  • Low sample consumption Low sample consumption
  • Under 2h bench time Under 2h bench time

The HTRF human GM-CSF kit is designed for the quantification of human GM-CSF release in cell supernatant.

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Overview

Also known as CSF2 (colony-stimulating factor 2), GM-CSF (granulocyte-macrophage colony-stimulating factor) is a hematopoietic growth factor and an immune modulator. Under immune stimuli, it is produced by T lymphocytes, macrophages, endothelial cells, and fibroblasts. It acts on precursor cells in the bone marrow to initiate their proliferation and differentiation into mature granulocytes and macrophages. GM-CSF stimulates multipotent progenitor cells.


Assessment of serum samples often requires enhanced sensitivity. In some cases, AlphaLISA assays may have sufficient sensitivity to enable the detection of low levels of analytes in serum or plasma. Check out the Human GM-CSF AlphaLISA kit’s analytical performances for more information or learn more about AlphaLISA. When assaying, always follow the recommended protocol and avoid highly haemolyzed samples. 

Benefits

  • SCALABLE FROM 96 TO 1536WELLS
  • NK CELL ACTIVATION READOUT

Assay principle

Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection by HTRF® reagents (384-well low-volume white plate or Cisbio low-volume 96-well plate in 20 µl). The antibodies labeled with the HTRF donor and acceptor are pre-mixed and added in a single dispensing step, to further streamline the assay procedure. The assay can be run up to a 1536-well format by simply resizing each addition volume proportionally.
GM-CSF kit assay principle

Assay data analysis

The 4 Parameter Logistic (4PL) curve is commonly recommended for fitting an ELISA standard curve. This regression enables the accurate measurement of an unknown sample across a wider range of concentrations than linear analysis, making it ideally suited to the analysis of biological systems like cytokine releases.

To fully understand how to deal with HTRF data processing and also 4PL 1/y² fitting, please visit this page.

Cisbio also worked with Myassays.com to help you in your data analysis. By clicking on this link you’ll be able to access a free online software to run your GM-CSF analysis.

Technical specifications of human GM-CSF kit

Sample size16 µL
Final assay volume20 µL
Kit componentsLyophilized standard, frozen detection antibodies, buffers & protocol.
LOD & LOQ (in Diluent)27 pg/mL & 47 pg/mL
Range47 – 4,500 pg/mL
Time to resultOvernight at RT
CalibrationNIBSC (09/136) value (IU/mL) = 0,07 x HTRF hGCSF HTRF
SpeciesHuman only

Intra and inter assay

Intra assay (n=24)

Sample Mean [GM-CSF] (pg/mL) CV
1 81 17%
2 2125 4%
3 6011 10%
Mean CV 10%

Inter assay (n=4)

Sample [GM-CSF] (pg/mL) Mean (delta ratio)
1 169 370 6%
2 774 1654 7%
3 4135 5313 6%
Mean CV 7%

GM-CSF secretion by PBMC co-stimulated with ionomycin and PMA

PBMCs, plated at 100,000 cells/well in a 96 well plate, were stimulated for 24h with 1µg/ml of ionomycin and with various concentrations of PMA. 16µl of supernatant were transferred into a 384 well (small volume) white detection plate to be analyzed by the Human GM-CSF Assay.
HTRF Cytokine cell-based assays

Monitor immune regulation the simple way - Flyers

Inflammation cell by cell

HTRF solutions for each cell type - Flyers

HTRF assays for Oncology and Inflammation

Signaling in the immune system - Brochures

Normalize HTRF cytokine assays with viability assay on Spectramax I3X-Minimax

In collaboration with Molecular Devices - Application Notes

HTRF measurement of cytokine release from fresh blood samples

In collaboration with Blood assay solution - Application Notes

Cytokine secretion profile on PBMCs

One sample to measure them all - Posters

Guidelines from PBMC isolation to cytokine assay optimisation

Perform and optimize cytokine assays on PBMC - Technical Notes

How to freeze PBMC

Guideline for PBMC freezing - Technical Notes

How to measure multiple cytokines from 1 sample

What to expect at the bench - Videos

Product Insert GM-CSF (h) Kit / 62HGMCSFPEG-62HGMCSFPEH

62HGMCSFPEG-62HGMCSFPEH - Product Insert

HTRF makes cytokine detection easy

A fun video introducing you to cytokines assays with HTRF - Videos

Cisbio Product Catalog 2019

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

Cytokine assays: a guide to success with HTRF

Setting up a successful cytokine assay - Guides

Portfolio of HTRF cytokine assays

This leaflet presents the analytical performances of each assay - Brochures

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

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