Human GM-CSF kit

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

No-wash
Low sample consumption
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.

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 size	16 µL
Final assay volume	20 µL
Kit components	Lyophilized standard, frozen detection antibodies, buffers & protocol.
LOD & LOQ (in Diluent)	27 pg/mL & 47 pg/mL
Range	47 – 4,500 pg/mL
Time to result	Overnight at RT
Calibration	NIBSC (09/136) value (IU/mL) = 0,07 x HTRF hGCSF HTRF
Species	Human 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.