The Human G-CSF kit is designed for the quantification of human G-CSF release in cell supernatant.
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  • No-wash
  • Highly specific
  • Under 2h bench time
The Human G-CSF kit is designed for the quantification of human G-CSF release in cell supernatant.
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In stock

Overview

Also known as CSF3 (colony-stimulating factor 3), G-CSF is a hematopoietic cytokine. It acts on precursor cells in the bone marrow to initiate their proliferation and differentiation into mature granulocytes, on hematopoietic stem cells to induce their mobilization in the blood stream, and on neurons to promote neurogenesis and block apoptosis. In myocardial infarction or ischemia, G-CSF promotes T cells' immune tolerance and tissue recovery. The absence of a functional G-CSF receptor is associated with congenital neutropenia.

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 G-CSF analysis.

Technical specifications of human G-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 [G-CSF] (pg/mL) CV
1 47 18%
2 2036 7%
3 3780 6%
Mean CV 10%

Each of the 3 samples was measured 24 times, and % CV was calculated for each sample.

Inter-assay (n=4)

Sample [G-CSF] (pg/mL) Mean (delta R) CV
1 87 222 13%
2 423 883 8%
3 2045 3161 6%
Mean CV 9%

Each of the samples was measured in 4 different experiments, and % CV was calculated for each sample.

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Guidelines from PBMC isolation to cytokine assay optimisation

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Product Insert G-CSF (h) Kit / 62HGCSFPEG-62HGCSFPEH

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