Recombinant Human GM-CSF, Animal-Free Protein

Animal-FreeTM Recombinant Human GM-CSF (Catalog # AFL215) stimulates cell proliferation of the TF-1 human erythroleukemic cell line. The ED50 for this effect is 6-30 pg/mL.

1 μg/lane of Animal-FreeTM Recombinant Human IL-4 (Catalog # AFL215) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 13 kDa.

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

Recombinant Human GM-CSF, Animal-Free Protein Summary

• Additional Information: Intended for preclinical researchers who may transition to GMP GM-CSF for their clinical work
• Details of Functionality: Measured in a cell proliferation assay using TF‑1 human erythroleukemic cells. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The ED₅₀ for this effect is 6-30 pg/mL.
  The specific activity of recombinant human GM-CSF is >1.0 x 10⁷ IU/mg, which is calibrated against the human GM-CSF WHO International Standard (NIBSC code: 88/646).
• Source: E. coli-derived human GM-CSF protein
  Ala18-Glu144
  Produced using non-animal reagents in an animal-free laboratory.
• Accession #: P04141.1
• N-terminal Sequence: Ala18
• Protein/Peptide Type: Animal-Free Recombinant Proteins
• Gene: CSF2
• Purity: >97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 14 kDa.
  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: 13 kDa, 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 GM-CSF, Animal-Free Protein

• colony stimulating factor 2 (granulocyte-macrophage)
• Colony-stimulating factor
• CSF
• CSF2
• CSF-2
• GMCSF
• GM-CSF
• GMCSFgranulocyte-macrophage colony-stimulating factor
• granulocyte-macrophage colony stimulating factor
• MGC131935
• MGC138897
• Molgramostim
• molgramostin
• Sargramostim

Background

GM-CSF was initially characterized as a factor that can support the in vitro colony formation of granulocyte-macrophage progenitors. It is also a growth factor for erythroid, megakaryocyte, and eosinophil progenitors. GM-CSF is produced by a number of different cell types (including T cells, B cells, macrophages, mast cells, endothelial cells, fibroblasts, and adipocytes) in response to cytokine or inflammatory stimuli. On mature hematopoietic cells, GM-CSF is a survival factor for and activates the effector functions of granulocytes, monocytes/macrophages, and eosinophils (1, 2). GM-CSF promotes a Th1 biased immune response, angiogenesis, allergic inflammation, and the development of autoimmunity (3 - 5). It shows clinical effectiveness in ameliorating chemotherapy-induced neutropenia, and GM-CSF transfected tumor cells are utilized as cancer vaccines (6, 7). The 22 kDa glycosylated GM-CSF, similar to IL-3 and IL-5, is a cytokine with a core of four bundled alpha -helices (8 - 12). Mature human GM-CSF shares 63%-70% amino acid sequence identity with canine, feline, porcine, and rat GM-CSF and 54% with mouse GM-CSF. GM-CSF exerts its biological effects through a heterodimeric receptor complex composed of GM-CSF R alpha /CD116 and the signal transducing common beta  chain (CD131) which is also a component of the high-affinity receptors for IL-3 and IL-5 (13, 14). In addition, GM-CSF binds a naturally occurring soluble form of GM-CSF R alpha (15). Human GM-CSF is active on canine and feline cells but not on murine cells (16-18).

1. Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
2. Barreda, D.R. et al. (2004) Dev. Comp. Immunol. 28:509.
3. Eksioglu, E.A. et al. (2007) Exp. Hematol. 35:1163.
4. Cao, Y. (2007) J. Clin. Invest. 117:2362.
5. Fleetwood, A.J. et al. (2005) Crit. Rev. Immunol. 25:405.
6. Heuser, M. et al. (2007) Semin. Hematol. 44:148.
7. Hege, K.M. et al. (2006) Int. Rev. Immunol. 25:321.
8. Kaushansky, K. et al. (1992) Biochemistry 31:1881.
9. Diederichs, K. et al. (1991) Science 254:1779.
10. Cantrell, M.A. et al. (1985) Proc. Natl. Acad. Sci. 82:6250.
11. Lee, F. et al. (1985) Proc. Natl. Acad. Sci. 82:4360.
12. Wong, G.G. et al. (1985) Science 228:810.
13. Onetto-Pothier, N. et al. (1990) Blood 75:59.
14. Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. 87:9655.
15. Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483.
16. Hogge, G.S. et al. (1990) Cancer Gene Ther. 6:26.
17. Sprague, W.S. et al. (2005) J. Comp. Pathol. 133:136.
18. Shanafelt, A.B. et al. (1991) J. Biol. Chem. 266:13804.

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

 

Bioinformatics

• Gene Symbol: CSF2
• Uniprot:
  • P04141.1()