< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
The Quantikine HS Human G-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure G-CSF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human G-CSF and antibodies raised against the recombinant protein. It has been shown to accurately quantitate recombinant human G-CSF. Results obtained using natural human G-CSF showed linear curves that were parallel to the standard curves obtained using the Quantikine HS kit standards. These results indicate that this kit can be used to determine relative mass values for natural human G-CSF.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Serum, EDTA Plasma, Heparin Plasma
Cell Culture Supernates
The recovery of G-CSF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples were spiked with high concentrations of G-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
G-CSF (granulocyte-colony stimulating factor) is a secreted glycoprotein that regulates the proliferation, differentiation, and activation of neutrophilic granulocyte lineage cells. It also enhances M-CSF induced monocyte differentiation and the proliferation of Th2-inducing dendritic cells. G-CSF promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia. It signals through G-CSF R/CD114 on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types. Mutations in G-CSF R are associated with congenital neutropenia.
Refer to the product for complete assay procedure.
Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal microplate shaker.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. PROTECT FROM LIGHT.
For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 30 minutes. For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 20 minutes.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.