Mouse G-CSF Quantikine ELISA Kit Summary Assay Type | Solid Phase Sandwich ELISA | Format | 96-well strip plate | Assay Length | 4.5 hours | Sample Type & Volume Required Per Well | Cell Culture Supernates (25 uL), Serum (50 uL) | Sensitivity | 5 pg/mL | Assay Range | 14.1 - 900 pg/mL (Cell Culture Supernates, Serum) | Specificity | Natural and recombinant mouse 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. |
Product Summary The Quantikine Mouse G-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse G-CSF in cell culture supernates and serum. It contains E. coli-expressed recombinant mouse G-CSF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse G-CSF accurately. Results obtained using natural mouse G-CSF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse G-CSF. Preparation and Storage Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. | Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
Background: G-CSFMouse granulocyte-colony stimulating factor (G-CSF) is a 24-25 kDa monomeric glycoprotein that regulates the proliferation, differentiation and activation of hematopoietic cells. Mouse G-CSF cDNA encodes a 208 amino acid (aa) precursor protein with a 30 aa signal sequence that is proteolytically cleaved to form a 178 aa O-glycosylated mature protein containing two intrachain disulfide bridges. In humans, two distinct cDNA clones, encoding a 204 aa form and a minor alternatively spliced 207 aa form of G-CSF precursors, have been isolated. Mouse and human G-CSF are 76% identical at the aa sequence level and the two proteins show species cross-reactivity. G-CSF is produced primarily by monocytes and macrophages upon activation by endotoxin, TNF-alpha or IL-1. Other cell types, including fibroblasts, endothelial cells, astrocytes and bone marrow stroma cells, can also secrete G-CSF after activation. In addition, various tumor cells express G-CSF constitutively. Mouse G-CSF receptor (G-CSF R) is a 120 kDa, type I transmembrane glycoprotein that belongs to the hematopoietin receptor superfamily. The mature protein is 812 amino acids in length and contains a 601 aa extracellular region, a 24 aa transmembrane segment, and a 187 aa cytoplasmic domain. The extracellular region contains multiple modules, including an N-terminal Ig-like domain, a cytokine receptor homology domain, and three fibronectin type III domains. Based on crystallographic study, G-CSF receptor forms a complex with the ligand in a 2:2 ratio. Mouse and human G-CSF receptor share 63% aa sequence identity. Cells known to express G-CSF R include monocytes and neutrophils, megakaryocytes and platelets, CD34+ CD33+ and CD34+ CD38+ hematopoietic progenitors, trophoblasts, endothelial cells and various tumor cell types. G-CSF is an important regulator for granulopoiesis in vivo. It has been demonstrated that G-CSF can support the growth of multi-lineage hematopoietic progenitor cells without influencing their commitment to the myeloid lineage and mobilize hematopoietic progenitor cells from the bone marrow into the bloodstream. On mature neutrophils, G-CSF may regulate neutrophil survival by controlling their rate of apoptosis. G-CSF has also been shown to enhance the functional capacity of mature neutrophils. As a consequence of its effects on hematopoietic progenitor cells, G-CSF has been shown to enhance monocytopoiesis in the presence M-CSF. Within the peripheral blood stem cell population mobilized with G-CSF, selective increases in the number of T helper 2-inducing dendritic cells are found. Long Name: | Granulocyte Colony Stimulating Factor | Entrez Gene IDs: | 1440 (Human); 12985 (Mouse) | Alternate Names: | C17orf33; chromosome 17 open reading frame 33; colony stimulating factor 3 (granulocyte); CSF3; CSF3OS; Filgrastim; GCSF; G-CSF; GCSFlenograstim; granulocyte colony-stimulating factor; Lenograstim; MGC45931; Pluripoietin |
Assay Procedure Refer to the product datasheet for the complete assay procedure. Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate. 1. Prepare all reagents, standard dilutions, and samples as directed in the product insert. 2. Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal. 3. Add 50 μL of Assay Diluent to each well. 4. Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours. 5. Aspirate each well and wash, repeating the process 4 times for a total of 5 washes. 6. Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours. 7. Aspirate and wash 5 times. 8. Add 100 μL Substrate Solution to each well. 9. Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm. |
