Recombinant Human GM-CSF, Animal-Free Protein

GMP Version Available: 215-GMP
GMP
Intended for preclinical researchers who may transition to GMP GM-CSF for their clinical work
Catalog # Availability Size / Price Qty
AFL215-01M
AFL215-200
AFL215-010
AFL215-050

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Recombinant Human GM-CSF, Animal-Free Protein Bioactivity
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Recombinant Human GM-CSF, Animal-Free Protein Summary

Animal Free Proteins
Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. Our products generated under animal-free conditions share the same biological activities as those produced using our standard laboratory techniques.

Product Specifications

Purity
>97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
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 #
N-terminal Sequence
Analysis
Ala18
Predicted Molecular Mass
14 kDa
SDS-PAGE
13 kDa, reducing conditions

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AFL215

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.

AFL215

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.

Animal Free Proteins
Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. Our products generated under animal-free conditions share the same biological activities as those produced using our standard laboratory techniques.

Scientific Data

Bioactivity Recombinant Human GM-CSF, Animal-Free Protein Bioactivity View Larger

Animal-FreeTMRecombinant 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.

SDS-PAGE Recombinant Human GM-CSF, Animal-Free Protein SDS-PAGE View Larger

1 μg/lane of Animal-FreeTMRecombinant 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GM-CSF

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).

References
  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.
Long Name
Granulocyte Macrophage Growth Factor
Entrez Gene IDs
1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)
Alternate Names
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

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

    FAQs

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