Human G-CSF Quantikine QuicKit ELISA

Name: Human G-CSF Quantikine QuicKit ELISA
Brand: R&D Systems
SKU: QK214

R&D Systems, Inc. a Bio-Techne Brand

Catalog # QK214

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Key Product Details

Assay Length:	80 minutes
Sample Type & Volume Required Per Well:	Cell Culture Supernates (25 uL), Serum (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
Sample Types:	Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Sensitivity:	3.57 pg/mL
Assay Range:	39.1 - 2,500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Human G-CSF Quantikine QuicKit ELISA Summary:	The Quantikine^® QuicKit™ Human G-CSF Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human G-CSF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human G-CSF and antibodies raised against the recombinant protein. Results obtained using natural human G-CSF showed linear curves that were parallel to the standard curves obtained using the QuicKit™ standards. These results indicate that this kit can be used to determine relative mass values for natural human G-CSF.

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Product Specifications for Human G-CSF Quantikine QuicKit ELISA

Measurement	Quantitative ELISA
Conjugate	HRP
Reactivity	Human
Specificity	Natural and recombinant human G-CSF.
Cross-reactivity	< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference	No significant interference observed with available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision

Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

	Intra-Assay Precision		Inter-Assay Precision
Sample	1	2	1	2
n	20	20	10	10
Mean (pg/mL)	260	1351	246	1426
Standard Deviation	3.36	26.2	27	70.9
CV%	1.3	1.9	11	5

Recovery

The recovery of human G-CSF spiked to three levels in samples throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Supernates (n=4)	101	93-105
EDTA Plasma (n=2)	102	100-106
Heparin Plasma (n=2)	94	90-99
Serum (n=2)	79	77-84

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human G-CSF were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.

Scientific Data Examples for Human G-CSF Quantikine QuicKit ELISA

	Human G-CSF ELISA Standard Curve
	Human G-CSF QuicKit Recovery Competitor Comparison G-CSF is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Culture media recovery is 102% compared to 122% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.
	Human G-CSF QuicKit Spiked Linearity Competitor Comparison G-CSF is spiked at high concentration in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity in serum is between 92%-112% compared to 121%-153% for the top competitor. The linearity in plasma is between 92%-93% compated to 143%-195% for the top competitor. Linearity is used to assess matrix effects and accurate measurement of sample values to provide confidence in your results.

Preparation and Storage

Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: G-CSF

Granulocyte-colony stimulating factor (G-CSF) is a 24-25 kDa monomeric glycoprotein that regulates the proliferation, differentiation, and activation of hematopoietic cells in the neutrophilic granulocyte lineage. Mature human G-CSF is a 178 amino acid (aa) O-glycosylated protein that contains two intrachain disulfide bridges. In humans, alternate splicing generates a second minor isoform with a 3 aa deletion. Mouse and human G-CSF share 76% aa sequence identity, and the two proteins show species cross-reactivity. G-CSF is produced by activated monocytes and macrophages, fibroblasts, endothelial cells, astrocytes, neurons, and bone marrow stroma cells. In addition, various tumor cells express G-CSF constitutively.

Human G-CSF receptor (G-CSF R) is a 120 kDa type I transmembrane glycoprotein that belongs to the hematopoietin receptor superfamily. The mature protein consists of a 603 aa extracellular domain (ECD), a 23 aa transmembrane segment, and a 186 aa cytoplasmic domain. The ECD contains an N-terminal Ig-like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Alternate splicing of human G-CSF R generates additional isoforms including a potentially soluble form of the receptor. The ECDs of mouse and human G-CSF R share 63% aa sequence identity. G-CSF R forms a complex with the ligand in a 2:2 ratio. It is expressed on monocytes, neutrophils, megakaryocytes, platelets, myeloid progenitors, trophoblasts and placenta, endothelial cells, and various tumor cell types.

G-CSF is an important regulator for granulopoiesis in vivo, and mutations in G-CSF R are associated with congenital neutropenia. G-CSF can support the growth of multilineage hematopoietic progenitor cells and mobilize them from the bone marrow into the bloodstream. G-CSF enhances the functional capacity of mature neutrophils and supports their survival by limiting the rate of apoptosis. G-CSF also enhances M-CSF induced monocytopoiesis from hematopoietic progenitor cells and stimulates the proliferation of peripheral Th2-inducing dendritic cells (30, 31). It promotes the development of T cell immune tolerance as well as tissue recovery following myocardial infarction and cerebral ischemia.

Item	Value
Long Name	Granulocyte Colony Stimulating Factor
Entrez Gene IDs	1440 (Human); 12985 (Mouse)
Alternate Names	C17orf33, CSF3, CSF3OS, Filgrastim, G-CSF, GCSF, GCSFlenograstim, Lenograstim, MGC45931, Pluripoietin, chromosome 17 open reading frame 33, colony stimulating factor 3 (granulocyte), granulocyte colony-stimulating factor

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