Description
Granulocyte macrophage colony stimulating factor (GM-CSF) is a potent species-specific stimulator of bone marrow cells. GM-CSF stimulates precursor cells of granulocytes, macrophages, and eosinophils. Human GM-CSF is a 14 - 17 kD multiple glycosylated protein containing 123 amino acid residues. Recombinant human GM-CSF (Cat. No. 550068) is supplied as a frozen liquid comprised of 0.22 µm sterile-filtered aqueous buffered solution containing bovine serum albumin with no preservatives. Recombinant human GM-CSF is ≥ 95% pure as determined by SDS-PAGE, and an absorbance assay based on the Beers-Lambert law. The endotoxin level is ≤ 0.1 ng per µg of human GM-CSF, as measured in a chromogenic LAL assay.
Preparation And Storage
Recommended Assay Procedures
Upon initial thawing, recombinant human GM-CSF (Cat. No. 550068) should be aliquoted into polypropylene microtubes and frozen at -80°C for future use. Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5-10 mg/mL carrier protein, such as human or bovine serum albumin, aliquoted and stored at -80°C. For in vitro biological assay use, carrier-protein concentrations of 0.5– 1 mg/mL are recommended. For use as an ELISA standard, carrier-protein concentrations of 5– 10 mg/mL are recommended. Failure to add carrier protein or store at indicated temperatures may result in a loss of activity. The product should not be diluted to less than 5 μg/mL for long term storage. Carrier proteins should be pre-screened for possible effects in each investigator's experimental system. Carrier proteins may have an undesired influence on experimental results due to toxicity, high endotoxin levels or possible blocking activity.
ELISA Standard: Recombinant human GM-CSF (Cat. No. 550068) can be useful as a quantitative standard for measuring human GM-CSF protein levels using sandwich ELISA with the purified BVD2-23B6 antibody (Cat. No. 554502) as a capture antibody and biotinylated BVD2-21C11 (Cat. No. 554505) as the detection antibody. To obtain linear standard curves, investigators may want to consider using doubling dilutions of recombinant human GM-CSF from 2,000 - 15 pg/mL to be included for each ELISA plate. For measuring human GM-CSF in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Human GM-CSF ELISA Set (Cat. No. 555126).
Bioassay: Investigators are advised that the Bioassay application is not routinely tested for this material and are highly encouraged to both titrate this material and include appropriate controls in relevant experiments. An activity range of 0.1 - 1.0 x 10^9 units/mg, encompassing an
ED50= 10 - 90 pg/mL, has previously been reported using TF-1 as indicator cells for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation.
Blocking: Recombinant human GM-CSF (Cat. No. 550068) can be useful as a blocking control for flow cytometric analysis when used with PE-conjugated BVD2-21C11 (Cat. No. 554507). Investigators are advised that the blocking application is not routinely tested for this material. Intracellular cytokine staining techniques and the use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Development References (4)
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Gasson JC. Molecular physiology of granulocyte-macrophage colony-stimulating factor. Blood. 1991; 77(6):1131-1145. (Biology). View Reference
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Kitamura T, Tange T, Terasawa T, et al. Establishment and characterization of a unique human cell line that proliferates dependently on GM-CSF, IL-3, or erythropoietin. J Cell Physiol. 1989; 140(2):323-334. (Biology). View Reference
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Miyatake S, Otsuka T, Yokota T, Lee F, Arai K. Structure of the chromosomal gene for granulocyte-macrophage colony stimulating factor: comparison of the mouse and human genes. EMBO J. 1985; 4(10):2561-2568. (Biology). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.