Recombinant Human M-CSF GMP Protein, CF

Catalog #: 216-GMP
3 Citations Datasheet / COA / SDS

Animal-Free

Catalog #	Availability	Size / Price	Qty
216-GMP-025
216-GMP-500

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Recombinant Human M-CSF GMP Protein Bioactivity

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Product Details

Citations (3)

Supplemental Products

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human M-CSF GMP Protein, CF Summary

Product Specifications

Purity

>97%, by SDS-PAGE with silver staining, under reducing conditions

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

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).

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.
Manufactured and tested under cGMP guidelines.

Accession #

NP_757350.2

N-terminal Sequence
Analysis

Met-Glu-Glu-Val-Ser-Glu-Try-(Cys)-Ser-His

Structure / Form

Disulfide-linked homodimer

Predicted Molecular Mass

18.5 kDa (monomer)

SDS-PAGE

37 kDa, non-reducing conditions

Product Datasheets

216-GMP

Product Datasheet

COA

SDS

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.

216-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 50-500 μg/mL in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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GMP-grade Recombinant Human M-CSF (Catalog # 216-GMP) stimulates cell proliferation in the M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line in a dose-dependent manner. The ED₅₀ for this effect is 0.5-1.5 ng/mL.

SDS-PAGE

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1 μg/lane of GMP-grade Recombinant Human M-CSF (Catalog # 216-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 18 kDa and 32 kDa, respectively.

Mass Spectrometry

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ESI analysis of GMP-grade Recombinant Human M-CSF (Catalog # 216-GMP). The peak at 37056 Da corresponds to the measured molecular weight of the intact dimer. The calculated mass for the monomer is 18535 Da.

Reconstitution Calculator

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.

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

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

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.

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.

Please read our complete Animal-Free Statement

Product Specific Notices

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

Related Research Areas

Citations for Recombinant Human M-CSF GMP Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Characterization of AB598, a CD39 Enzymatic Inhibitory Antibody for the Treatment of Solid Tumors
Authors: Anderson, AE;Parashar, K;Jin, K;Clor, J;Stagnaro, CE;Vani, U;Singh, J;Chen, A;Guan, Y;Talukdar, P;Sathishkumar, P;Juat, DJ;Singh, H;Kushwaha, R;Zhao, X;Kaplan, A;Seitz, L;Walters, MJ;Fernandez-Salas, E;Walker, NPC;Bowman, CE;
Molecular cancer therapeutics
Species: Human
Sample Types: Whole Cells
Dynamic prostate cancer transcriptome analysis delineates the trajectory to disease progression
Authors: M Bolis, D Bossi, A Vallerga, V Ceserani, M Cavalli, D Impellizzi, L Di Rito, E Zoni, S Mosole, AR Elia, A Rinaldi, R Pereira Me, E D'Antonio, M Ferrari, F Stoffel, F Jermini, S Gillessen, L Bubendorf, P Schraml, A Calcinotto, E Corey, H Moch, M Spahn, G Thalmann, M Kruithof-d, MA Rubin, JP Theurillat
Nature Communications, 2021-12-02;12(1):7033.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
Development, functional characterization and validation of methodology for GMP-compliant manufacture of phagocytic macrophages: A novel cellular therapeutic for liver cirrhosis.
Authors: Fraser A, Pass C, Burgoyne P, Atkinson A, Bailey L, Laurie A, W A McGowan N, Hamid A, Moore J, Dwyer B, Turner M, Forbes S, Campbell J
Cytotherapy, 2017-06-30;19(9):1113-1124.
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture