| Field | Specification |
|---|---|
| Target | |
| Alternative names | Granulocyte-macrophage colony-stimulating factor;GM-CSF;Colony-stimulating factor;CSF;Csf2;Csfgm; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: A18-K141 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies mouse granulocyte-macrophage colony-stimulating factor (gene Csf2) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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.
GM-CSF (CSF2) is a cytokine that drives the growth and differentiation of hematopoietic progenitors into granulocytes, macrophages and other myeloid cells. Produced by T cells, macrophages, endothelial cells and fibroblasts, it also activates mature myeloid cells, so it is a frequent target in studies of inflammation, autoimmunity and dendritic cell culture.
Kit Components
| Anti-Mouse Csf2 Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Csf2 Standard | 2 vials, 10 ng/tube |
| Mouse Csf2 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 | Mouse |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 15.6 pg/mL–1,000 pg/mL |
| Sensitivity | <1 pg/mL |
| Standard | Recombinant mouse Csf2 (Ala18–Lys141), expressed in E. coli |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 4.1–7.0% (3 samples, n = 16 each) |
| Inter-assay CV | 4.4–7.0% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:1. In the supplier’s tests, serum and plasma samples measured below the lowest standard.
Example standard curve (OD450, pg/mL): 0 = 0.033; 15.6 = 0.096; 31.2 = 0.155; 62.5 = 0.265; 125 = 0.458; 250 = 0.83; 500 = 1.362; 1000 = 2.055. 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 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.016 | 0.118 | 0.232 | 0.425 | 0.797 | 1.329 | 1.883 | 2.222 |
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) | 32 | 234 | 480 | 35 | 213 | 448 |
| Standard deviation | 1.34 | 17.8 | 19.68 | 1.54 | 14.91 | 21.05 |
| CV (%) | 4.2% | 7% | 4.1% | 4.4% | 7% | 4.7% |
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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