| Field | Specification |
|---|---|
| Target | |
| Alternative names | Granulocyte colony-stimulating factor;G-CSF;Pluripoietin;Filgrastim;Lenograstim;CSF3;C17orf33, GCSF; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: T31-P204 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies human granulocyte colony-stimulating factor (gene CSF3) in cell culture supernatants, serum and plasma (heparin, EDTA, citrate). Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
G-CSF (granulocyte colony-stimulating factor, gene CSF3) is the main cytokine that drives the production, maturation and release of neutrophils from the bone marrow. It is measured in studies of infection, inflammation and bone marrow recovery, and recombinant G-CSF is used clinically to treat neutropenia.
Kit Components
| Anti-Human CSF3 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Human CSF3 Standard | 2 vials, 10 ng/tube |
| Human CSF3 Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Human |
|---|---|
| Sample types | Cell culture supernatants, serum and plasma (heparin, EDTA, citrate) |
| Assay range | 31.2 pg/mL–2,000 pg/mL |
| Sensitivity | <4 pg/mL |
| Standard | Recombinant human CSF3 (Thr31–Pro204), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.5–6.4% (3 samples, n = 16 each) |
| Inter-assay CV | 6.2–7.9% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Example standard curve (OD450, pg/mL): 0 = 0.088; 31.2 = 0.15; 62.5 = 0.199; 125 = 0.307; 250 = 0.484; 500 = 0.829; 1000 = 1.454; 2000 = 2.558. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (pg/ml) | 0 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 | 2000 |
| O.D. | 0.088 | 0.15 | 0.199 | 0.307 | 0.484 | 0.829 | 1.454 | 2.558 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 62 | 237 | 1001 | 64 | 230 | 966 |
| Standard deviation | 3.96 | 16.26 | 45.04 | 4.99 | 18.17 | 59.89 |
| CV (%) | 6.4% | 6.4% | 4.5% | 7.8% | 7.9% | 6.2% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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Jinlan Dong et al. (2023) Extracellular proteins as potential biomarkers in Sepsis-related cerebral injury. Frontiers in Immunology. 10.3389/fimmu.2023.1128476
Sobrino Tomás et al. (2019) Association of High Serum Levels of Growth Factors with Good Outcome in Ischemic Stroke: a Multicenter Study. Translational Stroke Research. 10.1007/s12975-019-00747-2
Qiao Shan et al. (2023) Abnormal DNA methylation analysis of leucine-rich glioma-inactivated 1 antibody encephalitis reveals novel methylation-driven genes related to prognostic and clinical features. Clinical Epigenetics. 10.1186/s13148-023-01550-5
Kewen Ding et al. (2021) Effect of steam-processing of the Panax ginseng root on its inducible activity on granulocyte-colony stimulating factor secretion in intestinal epithelial cells in vitro. Journal of Ethnopharmacology. 10.1016/j.jep.2021.114927
Smaranika Rahman et al. (2022) Increased serum resistin but not G-CSF levels are associated in the pathophysiology of major depressive disorder: Findings from a case-control study. PLoS One. 10.1371/journal.pone.0264404
Elena López-Cancio et al. (2016) Reported Prestroke Physical Activity Is Associated with Vascular Endothelial Growth Factor Expression and Good Outcomes after Stroke. Journal of Stroke & Cerebrovascular Diseases. 10.1016/j.jstrokecerebrovasdis.2016.10.004
Danyang Wang et al. (2017) Improving transcriptional activity of human cytomegalovirus major immediate-early promoter by mutating NF-κB binding sites. Protein Expression and Purification. 10.1016/j.pep.2017.09.008
