Human G-CSF Quantikine QuicKit ELISA
R&D Systems, part of Bio-Techne | Catalog # QK214
Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (25 µL), Serum (25 µL), EDTA Plasma (25 µL), Heparin Plasma (25 µL)
Sensitivity
3.57 pg/mL
Assay Range
39.1-2500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Human G-CSF Quantikine QuicKit ELISA
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.
Product Specifications
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 - Ten 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, 39.1 pg/mL.
ND=Non-detectable
Cell Culture Supernates - Human peripheral blood mononuclear cells (PBMCs)
(1 x 106
cells/mL) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum,
2 mM L-glutamine, 100 U/mL penicillin, and 100 ug/mL streptomycin sulfate. Cells were left
unstimulated or stimulated with 10 ug/mL PHA for 1 or 5 days. Aliquots of the cell culture
supernates were removed and assayed for levels of human G-CSF.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | ND | ND |
| Stimulated | 915 | 1184 |
THP-1 cells (2.5 x 105
cells/mL) were cultured in RPMI 1640 supplemented with 10% fetal bovine
serum, 2 mM L-glutamine, 100 U/mL penicillin, 100 µg/mL streptomycin sulfate, and
50 µM beta-mercaptoethanol. To differentiate the cells into macrophages, 50 ng/mL PMA was
added when seeding cells and incubated overnight. The next day, media was removed, and
cells were washed once with media before replacing normal media and incubated for 2 days.
Cells were then left unstimulated or stimulated with 100 ng/mL or 1000 ng/mL LPS for 1 day.
Aliquots of the cell culture supernates were removed and assayed for levels of human G-CSF.
| Condition | Day 1 (pg/mL) |
| Unstimulated | ND |
| Stimluated with 100 ng/mL LPS | 1813 |
| Stimluated with 1000 ng/mL LPS | 2721 |
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, EDTA Plasma, Heparin Plasma, Serum
| 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 | 27.0 | 70.9 |
| CV% | 1.3 | 1.9 | 11.0 | 5.0 |
Recovery for Human G-CSF Quantikine QuicKit ELISA
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 Images 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
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 QuicKit ELISA
Product Specific Notices for Human G-CSF Quantikine QuicKit ELISA
For research use only
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