Mouse Macrophage Colony stimulating Factor Recombinant

Description

Methods (54)


accession	P07141

Source	Optimized DNA sequence encoding mouse M-CSF extracellular domain was expressed in Escherichia Coli.
Molecular weight	Mouse MCSF, generated by the proteolytic removal of the signal peptide and propeptide, The molecule has a calculated molecular mass of approximately 18 kDa. Recombinant Mouse M-CSF is a disulfide-linked homodimeric protein consisting of two 156 amino acid residue subunits, andmigrates as an approximately 37 kDa protein under non-reducing and as 18-19kDa under reducing conditions in SDS-PAGE.
Purity	>95%, as determined by SDS-PAGE and HPLC
Biological Activity	The ED(50) was determined by the dose-dependent stimulation of the proliferation of of M-NFS-60 cells is < 1.0 ng/ml, corresponding to a specific activity of > 1 x 106 units/mg.
Protein Sequence
Endotoxin	Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Presentation	Recombinant mouse MCSF was lyophilized from a 0.2 μm filtered PBS solution.
Reconstitution	A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
Storage	The lyophilized protein is stable for at least 2 years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage	This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.

Interactor	P03228
Interactor	P03228
Interactor	P09581 CSF1R_MOUSE
Interactor	P46414
Interactor	P07333 CSF1R_HUMAN
Biological Process	Immunity
Biological Process	Inflammatory-response
Molecular function	Cytokine
Molecular function	Growth-factor

Methods


Lung-derived prominin-1. Exposure of prominin-1+ cells to M-CSF for 7 days resulted in formation of E.coli phagocytising F4/80-positive cells .
Osteoblast, osteoclast and macrophage differentiation. Bone marrow derived monocytes were seeded in 24-well culture plates and induced to differentiate into macrophages or osteoclasts via the addition of M-CSF or M-CSF + RANKL respectively.
Cell activation assay and immunodetection of cytokines Single spleen cell suspensions were prepared by mechanical disruption on a cell strainer (70 µm pore diameter , , ) in Dulbecco's D-PBS . Red blood cells were lysed using Hemolytic Gey's Solution as previously described
Osteoclastogenesis assays Osteoclastogenesis assays were performed as in ⁵ cells were plated in each well of a 24-well dish with 60 ng/mL RANKL for 3 days. After 3 days, 2×10³ MDA-231, shControl, or shEGFR-MDA-231 cells were plated with the… Osteoclastogenesis assays were performed as in ⁵ cells were plated in each well of a 24-well dish with 60 ng/mL RANKL for 3 days. After 3 days, 2×10³ MDA-231, shControl, or shEGFR-MDA-231 cells were plated with the bone marrow cells, with 60 ng/mL RANKL and 10 ng/mL MCSF and grown for 3 days before TRAP staining. For TRAP staining, cells were washed with 1×PBS, fixed with ice cold methanol for 10 minutes, and stained in fresh TRAP solution for 15 minutes at 37°C. TRAP solution was replaced with 1×PBS for cell counting under the microscope. Read more
PLCγ2 is required for in vitro osteoclast development. (a) Representative TRAP-stained images of wild-type and PLCγ2−/− bone marrow cells cultured in the presence of the indicated concentrations of recombinant murine M-CSF and RANKL for 4 days.
Generation of conditional TAK1-deficient mice. Colony formation by BM cells from Map3k7
Generation of bone marrow-derived macrophages Bone marrow-derived macrophages (BMDM) were generated by flushing the bone marrow from the femurs of BALB/c mice with RPMI 1640 media. Freshly collected bone marrow cells were cultured overnight in complete media (CM; Bone marrow-derived macrophages (BMDM) were generated by flushing the bone marrow from the femurs of BALB/c mice with RPMI 1640 media. Freshly collected bone marrow cells were cultured overnight in complete media (CM; RPMI 1640, 10% fetal bovine serum [Atlanta, ], 10 mM HEPES buffer, 10 mM nonessential amino acids, 10 mM sodium pyruvate) containing 5 ng/ml recombinant murine M-CSF . The non-adherent cells were then collected and cultured for an additional six days in CM with 30 ng/ml M-CSF to generate macrophages. Read more
Pro-resorptive cytokine expression and osteoclastogenesis in the synovial fluid of patients with gouty arthritis. PBMCs, SFMCs, and T cell-depleted SFMCs (3 × 105 cells/well) were cultured for 10 days in the presence of macrophage colony-stimulating factor (M-CSF; 30 ng/ml) at the different RANKL concentrations and then were stained for tartrate-resistant acid… PBMCs, SFMCs, and T cell-depleted SFMCs (3 × 105 cells/well) were cultured for 10 days in the presence of macrophage colony-stimulating factor (M-CSF; 30 ng/ml) at the different RANKL concentrations and then were stained for tartrate-resistant acid phosphatase . Read more
TcREG inhibit osteoclast activity in an antigen- and contact-independent manner by secreted cytokines. Cells were re-fed with medium containing M-CSF and RANKL every 2 to 3 days.
Induction of 2nd iPSCs All isolated somatic cells were cultured in the presence of Dox (2 ug/ml) for induction of 2^nd iPSCs. Fibroblasts and hematopoietic cells were cultured in ES/iPSC medium. FLCD45 were cultured in the presence of 10… All isolated somatic cells were cultured in the presence of Dox (2 ug/ml) for induction of 2^nd iPSCs. Fibroblasts and hematopoietic cells were cultured in ES/iPSC medium. FLCD45 were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse EPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6, 10 ng/ml mouse Flt3 ligand, 10 ng/ml mouse GM-CSF, 10 ng/ml mouse VEGF and 50 ng/ml mouse SCF . HSCs, HPCs and MPs were cultured in the presence of 10 ng/ml human TPO, 10 ng/ml mouse IL-3, 10 ng/ml mouse IL-6 and 10 ng/ml mouse Flt3 ligand . Macrophages were cultured in the presence of 5 ng/ml M-CSF . Read more

Mouse Macrophage Colony stimulating Factor Recombinant

Mouse Macrophage Colony stimulating Factor Recombinant

Description

Methods (54)

Methods

Lung-derived prominin-1.

Osteoblast, osteoclast and macrophage differentiation.

Cell activation assay and immunodetection of cytokines

Osteoclastogenesis assays

PLCγ2 is required for in vitro osteoclast development.

Generation of conditional TAK1-deficient mice.

Generation of bone marrow-derived macrophages

Pro-resorptive cytokine expression and osteoclastogenesis in the synovial fluid of patients with gouty arthritis.

TcREG inhibit osteoclast activity in an antigen- and contact-independent manner by secreted cytokines.

Induction of 2nd iPSCs

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