Recombinant Human GM-CSF, Animal-Free Protein

Name: Recombinant Human GM-CSF, Animal-Free Protein
Brand: R&D Systems
SKU: AFL215

Catalog # AFL215 | R&D Systems, Inc. a Bio-Techne Brand

Product Datasheet / COA / SDS

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

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Setting the standard in quality research reagents

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Source:	E. coli
Accession #:	P04141.1
Applications:	Bioactivity

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Technical Data
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Product Specifications for Recombinant Human GM-CSF, Animal-Free Protein

Source	E. coli-derived human GM-CSF protein Ala18-Glu144 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	<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Ala18
Predicted Molecular Mass	14 kDa
SDS-PAGE	14 kDa
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).

Scientific Data Examples for Recombinant Human GM-CSF, Animal-Free Protein

	Recombinant Human GM-CSF, Animal-Free Protein Bioactivity Animal-Free^TMRecombinant 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.
	Recombinant Human GM-CSF, Animal-Free Protein SDS-PAGE 1 μg/lane of Animal-Free^TMRecombinant 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.

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.

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.

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).

Item	Value
References	Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653. Barreda, D.R. et al. (2004) Dev. Comp. Immunol. 28:509. Eksioglu, E.A. et al. (2007) Exp. Hematol. 35:1163. Cao, Y. (2007) J. Clin. Invest. 117:2362. Fleetwood, A.J. et al. (2005) Crit. Rev. Immunol. 25:405. Heuser, M. et al. (2007) Semin. Hematol. 44:148. Hege, K.M. et al. (2006) Int. Rev. Immunol. 25:321. Kaushansky, K. et al. (1992) Biochemistry 31:1881. Diederichs, K. et al. (1991) Science 254:1779. Cantrell, M.A. et al. (1985) Proc. Natl. Acad. Sci. 82:6250. Lee, F. et al. (1985) Proc. Natl. Acad. Sci. 82:4360. Wong, G.G. et al. (1985) Science 228:810. Onetto-Pothier, N. et al. (1990) Blood 75:59. Hayashida, K. et al. (1990) Proc. Natl. Acad. Sci. 87:9655. Pelley, J.L. et al. (2007) Exp. Hematol. 35:1483. Hogge, G.S. et al. (1990) Cancer Gene Ther. 6:26. Sprague, W.S. et al. (2005) J. Comp. Pathol. 133:136. 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	CSF, CSF-2, CSF2, Colony-stimulating factor, GM-CSF, GMCSF, GMCSFgranulocyte-macrophage colony-stimulating factor, MGC131935, MGC138897, Molgramostim, Sargramostim, colony stimulating factor 2 (granulocyte-macrophage), granulocyte-macrophage colony stimulating factor, molgramostin

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 GM-CSF, Animal-Free Protein

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Recombinant Human GM-CSF, Animal-Free Protein

Catalog # AFL215 | R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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

Scientific Data Examples for Recombinant Human GM-CSF, Animal-Free Protein

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: GM-CSF

Manufacturing Specifications

Citations for Recombinant Human GM-CSF, Animal-Free Protein

Product Documents for Recombinant Human GM-CSF, Animal-Free Protein

Product Datasheets

COA

Certificate of Analysis

SDS

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