Human G-CSF Quantikine ELISA Kit
R&D Systems, part of Bio-Techne | Catalog # DCS50
Key Product Details
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)
Sensitivity
20 pg/mL
Assay Range
39-2500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Product Summary for Human G-CSF Quantikine ELISA Kit
The Quantikine Human G-CSF Immunoassay is a 3.5 or 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 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 kit standards. These results indicate that this kit can be used to determine relative mass values for natural human G-CSF.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
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.
Sample Values
Serum/Plasma - Thirty-seven serum and plasma samples from apparently healthy volunteers
were evaluated for the presence of human G-CSF in this assay. No medical histories were
available for the donors used in this study. All samples measured less than the lowest human
G-CSF standard, 39 pg/mL.
Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106
cells/mL) were
cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta-mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and stimulated
with 10 μg/mL PHA. Aliquots of the culture supernate were removed on days 1 and 5 and
assayed for levels of natural human G-CSF.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 258 | 118 |
| Stimulated | 854 | 1139 |
Precision
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.
Cell Culture Supernates
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 177 | 515 | 1047 | 143 | 818 | 1514 |
| Standard Deviation | 4.5 | 9.4 | 22.0 | 17.0 | 72.0 | 77.1 |
| CV% | 2.5 | 1.8 | 2.1 | 11.9 | 8.8 | 5.1 |
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 280 | 827 | 1696 | 176 | 1094 | 2169 |
| Standard Deviation | 7.8 | 14.1 | 19.5 | 7.3 | 34.6 | 81.6 |
| CV% | 2.8 | 1.7 | 1.1 | 4.1 | 3.2 | 3.8 |
Recovery for Human G-CSF Quantikine ELISA Kit
The recovery of G-CSF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 94 | 85-110 |
| Citrate Plasma (n=5) | 90 | 78-111 |
| EDTA Plasma (n=5) | 93 | 80-117 |
| Heparin Plasma (n=5) | 87 | 76-103 |
| Serum (n=5) | 88 | 75-106 |
Linearity
To assess the linearity of the assay, five 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.
Scientific Data Images for Human G-CSF Quantikine ELISA Kit
Human G-CSF ELISA Cell Culture Supernate Standard Curve
Human G-CSF ELISA Serum/Plasma Standard Curve
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
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.
Long Name
Granulocyte Colony Stimulating Factor
Alternate Names
C17orf33, CSF3, CSF3OS, Filgrastim, GCSF, Lenograstim, Pluripoietin
Gene Symbol
CSF3
Additional G-CSF Products
Product Documents for Human G-CSF Quantikine ELISA Kit
Product Specific Notices for Human G-CSF Quantikine ELISA Kit
For research use only
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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.