Human GM-CSF Assay Kit

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.

Cisbio's human GM-CSF kit is a homogeneous, add and read assay to measure G-CSF secretion in cell supernatant. It offers a fast and efficient alternative to ELISA.

Sample size 16 µL
Final assay volume 20 µL
Kit components Lyophilized standard, frozen detection antibodies, buffers & protocol
LOD & LOQ (in diluent) 29 pg/mL & 54 pg/mL
Range 54 – 8,500 pg/mL
Time to result 3h @ RT
Calibration NIBSC (88/646) value (IU/mL) = 0.01 x HTRF hGM-CSF value (pg/mL)
Species Human and cynomolgus
 
Download Insider tips for PBMC isolation Technical Note
Download cytokine release from fresh blood samples application notes

Assay principle

The assay protocol, using a Cisbio 96-well low volume or a 384-well small volume white plate (20 µL final), is described below. Cell supernatant, sample or standard are dispensed directly into the assay plate for the detection by HTRF® reagents. 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 in 96 to 1536-well plates by simply resizing each addition volume proportionally.

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.

Add

Cytokines assay principle add

Read

Cytokines assay principle read

(1) Assay data analysis:

  • "Guidelines for data reduction" button will show you how to deal with HTRF data processing and 4PL 1/y2 fitting.
  • "MyAssays" button will direct you to a free online software able to run IFNγ analysis.

 

How to use HTRF cytokine assays

How to measure multiple cytokines from 1 sample

Analytical performance

Precision

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%

Assay validation

GM-CSF secretion by PBMCs 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 (SV) white detection plate to be analyzed by the Human GM-CSF Assay Kit.

Part#, inserts & MSDS

Ordering Info

DescriptionCat. noProduct insertMSDS
Human GM-CSF kit - 500 tests62HGMCSFPEG
Human GM-CSF kit - 10,000 tests62HGMCSFPEH
-

Companion products

DescriptionCat. noProduct insertMSDS
Human GM-CSF kit - 3 Standards62HGMCSFCD3
-
Human GM-CSF kit - Standard62HGMCSFCDA
-

Documents