Recombinant Human M-CSF, Animal-Free Protein

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human M-CSF, Animal-Free Protein Summary

• Additional Information: Intended for preclinical researchers who may transition to GMP M-CSF for their clinical work
• Details of Functionality: Measured in a cell proliferation assay using M‑NFS‑60 mouse myelogenous leukemia lymphoblast cells. Nakoinz, I. et al. (1990) J. Immunol. 145:860. The ED₅₀ for this effect is 0.5-1.5 ng/mL.
  The specific activity of Recombinant Human M-CSF is >6.0 x 10⁷ IU/mg, which is calibrated against the human M-CSF WHO International Standard (NIBSC code: 89/512).
• Source: E. coli-derived human M-CSF protein
  Glu33-Ser190, with an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
• Accession #: NP_757350.2
• N-terminal Sequence: Met
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Gene: CSF1
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 18.5 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 37 kDa, non-reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human M-CSF, Animal-Free Protein

• colony stimulating factor 1 (macrophage)
• CSF1
• CSF-1
• Lanimostim
• macrophage colony stimulating factor
• macrophage colony-stimulating factor 1
• MCSF
• M-CSF
• MCSFlanimostim
• MGC31930

Background

M-CSF, also known as CSF-1, is a four-alpha -helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation (1 - 3). M-CSF is also essential for the survival and proliferation of osteoclast progenitors (1, 4). M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis (2, 3). M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta (5). Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells (1 - 5). The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur (3 - 9). Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen (8). Shorter transcripts encode M-CSF that lacks cleavage and PG sites and produces an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer (7). Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor (10, 11). The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively (12, 13). Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.

1. Pixley, F.J. and E.R. Stanley (2004) Trends Cell Biol. 14:628.
2. Chitu, V. and E.R. Stanley (2006) Curr. Opin. Immunol. 18:39.
3. Fixe, P. and V. Praloran (1997) Eur. Cytokine Netw. 8:125.
4. Ryan, G.R. et al. (2001) Blood 98:74.
5. Makrigiannakis, A. et al. (2006) Trends Endocrinol. Metab. 17:178.
6. Nandi, S. et al. (2006) Blood 107:786.
7. Rettenmier, C.W. and M.F. Roussel (1988) Mol. Cell Biol. 8:5026.
8. Suzu, S. et al. (1992) J. Biol. Chem. 267:16812.
9. Manos, M.M. (1988) Mol. Cell. Biol. 8:5035.
10. Koths, K. (1997) Mol. Reprod. Dev. 46:31.
11. Jang, M-H. et al. (2006) J. Immunol. 177:4055.
12. Kawasaki, E.S. et al. (1985) Science 230: 291.
13. Wong, G.G. et al. (1987) Science 235:1504.

Manufacturing Process

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

• All molecular biology procedures use animal-free media and dedicated labware.
• Dedicated fermentors are utilized in committed animal-free areas.

Purification

• Protein purification columns are animal-free.
• Bulk proteins are filtered using animal-free filters.
• Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

• Low Endotoxin Level.
• No impairment of biological activity.
• High quality product obtained under stringent conditions.
• For ex vivo research or bioproduction, additional documentation can be provided.

Please read our complete Animal-Free Statement

 

Publications for M-CSF (AFL216)(1)

Publication using AFL216

• JA Hutchinson, N Ahrens, EK Geissler MITAP-compliant Characterisation of Human Regulatory Macrophages Transpl. Int., 2017-06-26;0(0):. 2017-06-26 [PMID: 28543878] (Bioassay, Human)

Bioinformatics

• Gene Symbol: CSF1
• Uniprot:
  • NP_757350.2()