Recombinant Human M-CSF, Animal-Free Protein

Name: Recombinant Human M-CSF, Animal-Free Protein
Brand: R&D Systems
SKU: AFL216

Catalog # AFL216 | R&D Systems, Inc. a Bio-Techne Brand

Citations (1)

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Intended for preclinical researchers who may transition to GMP M-CSF for their clinical work

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Learn more about Animal-Free Recombinant Proteins

Source:	E. coli
Accession #:	NP_757350.2
Structure / Form:	Disulfide-linked homodimer
Applications:	Bioactivity

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Technical Data
Citations (1) / Reviews

Product Specifications for Recombinant Human M-CSF, Animal-Free Protein

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.
Purity	>97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin level	<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Met
Predicted Molecular Mass	18.5 kDa (monomer)
SDS-PAGE	18.5 kDa (monomer)
Activity	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).

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AFL216

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

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Background: M-CSF

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.

Item	Value
References	Pixley, F.J. and E.R. Stanley (2004) Trends Cell Biol. 14:628. Chitu, V. and E.R. Stanley (2006) Curr. Opin. Immunol. 18:39. Fixe, P. and V. Praloran (1997) Eur. Cytokine Netw. 8:125. Ryan, G.R. et al. (2001) Blood 98:74. Makrigiannakis, A. et al. (2006) Trends Endocrinol. Metab. 17:178. Nandi, S. et al. (2006) Blood 107:786. Rettenmier, C.W. and M.F. Roussel (1988) Mol. Cell Biol. 8:5026. Suzu, S. et al. (1992) J. Biol. Chem. 267:16812. Manos, M.M. (1988) Mol. Cell. Biol. 8:5035. Koths, K. (1997) Mol. Reprod. Dev. 46:31. Jang, M-H. et al. (2006) J. Immunol. 177:4055. Kawasaki, E.S. et al. (1985) Science 230: 291. Wong, G.G. et al. (1987) Science 235:1504.
Long Name	Macrophage Colony Stimulating Factor
Entrez Gene IDs	1435 (Human); 12977 (Mouse)
Alternate Names	CSF-1, CSF1, Lanimostim, M-CSF, MCSF, MCSFlanimostim, MGC31930, colony stimulating factor 1 (macrophage), macrophage colony stimulating factor, macrophage colony-stimulating factor 1

Manufacturing Specifications

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

Citations for Recombinant Human M-CSF, Animal-Free Protein (1)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human M-CSF, Animal-Free Protein include:

Species: Human
Sample Types: Whole Cells
Applications: Bioassay

JA Hutchinson et al. (2017-06-26), MITAP-compliant Characterisation of Human Regulatory Macrophages Transpl. Int., 2017-00(0).
PMID: 28543878

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