Recombinant Mouse M-CSF Protein (CHO)

Biological Activity

Macrophage colony stimulating factor (M-CSF), also known as colony stimulating factor-1 (CSF-1), is a hematopoietic growth factor. It can stimulate the survival, proliferation and differentiation of mononuclear phagocytes, and can stimulate the proliferation and movement of macrophages. M-csf is mainly produced by monocytes, macrophages, fibroblasts and endothelial cells. The interaction of M-CSF with its receptor C-FMS is associated with growth, invasion, and metastasis in a variety of diseases, including breast cancer and endometrial cancer.

Measured in a cell proliferation assay using Murine M-NFS-60 cells, the ED50 is < 3.0 ng/ml, corresponding to a specific activity of > 3.3 × 10⁵ units/mg.

Accession # P07141

Protein Construction: M-CSF (Lys33-Pro187)

Endotoxin: < 0.2 EU/μg

Apparent Molecular Weight: 35~44 kDa

Lyophilized after extensive dialysis against PBS.

BMDMs were isolated from adult (8–10 week old) female C57BL/6 mice. After euthanasia, femurs were removed and stripped of muscle, exposed bone was washed in 70 % EtOH and rinsed with RPMI media (with 1 % penicillin/streptomycin) (RPMI-PS). Bone marrow cells were flushed from bone shafts into fresh RPMI media with 1 % penicillin/streptomycin and 10 % FBS (cRPMI) and then manually separated with two 23-gauge needles. Cells were pelleted by centrifugation at 300 g for 7 min and 4 °C, and the cell pellet was resuspended in 5 ml red blood cell lysis buffer (M58116). Next, RPMI-PS was added to the cell solution and centrifugation repeated. The cell pellet was washed twice with RPMI, resuspended in cRPMI and plated at a density of 2 × 10⁷ cells/dish in 60 ml of cRPMI media supplemented with 20 ng/ml of M-CSF (M11486). After 3 days, half the volume of media was replaced. After four additional incubation days, cells were washed with PBS and 3 ml of non-enzymic cell dissociation buffer was added. Following 10 min at 37 °C and 5 % CO2, cells were removed, centrifuged at 300 g for 7 min at 4 °C and then plated for infection or lipopolysaccharide (LPS) stimulation.

Chemical Information

Form	Lyophilized powder
Solubility (25°C)	Reconstitute the lyophilized powder in ddH2O or PBS up to 100 μg/ml.
Storage	Stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days and at -20°C for 3 months.

References

[1] Jiawei Ji, et al. Int Immunopharmacol. M-CSF and prostratin induced Mregs promote immune tolerance in transplanted mice through Arg-1 pathway

[2] Carolina Duarte, et al. Biochem Biophys Res Commun. Age-dependent effects of the recombinant spike protein/SARS-CoV-2 on the M-CSF- and IL-34-differentiated macrophages in vitro

[3] Chia-Sing Lu, et al. J Hematol Oncol. Oct4 promotes M2 macrophage polarization through upregulation of macrophage colony-stimulating factor in lung cancer

[4] Lauar de Brito Monteiro, et al. Immunobiology. M-CSF- and L929-derived macrophages present distinct metabolic profiles with similar inflammatory outcomes

[5] B D Chen. Blood. In vivo administration of recombinant human interleukin-1 and macrophage colony-stimulating factor (M-CSF) induce a rapid loss of M-CSF receptors in mouse bone marrow cells and peritoneal macrophages: effect of administration route