Granulocyte-Macrophage Colony Stimulating Factor is a 22 kD, pleiotropic cytokine that is a white blood cell growth factor. It controls the production and function of blood cells by stimulating stem cells to produce granulocytes and monocytes. GM-CSF differs from G-CSF in that it affects more cell types including macrophages and eosinophils. Moreover, GM-CSF is part of the immune/inflammatory cascade, a process crucial for fighting infection. Interestingly, GM-CSF expression may have pathological implications. Autocrine expression of GM-CSF in myeloid leukemia cells is suspected to play a role in neoplasia, the formation of a new and abnormal growth of tissue. Additionally, GM-CSF expression has also been documented in certain solid tumors. There have also been reports of GM-CSF in synovial fluid from patients with arthritis suggesting that GM-CSF may play a role in tissue damage associated with the inflammatory process. Blocking GM-CSF is thought to have therapeutic potential by reducing inflammation. Some drugs are currently being developed to block GM-CSF.
The predicted molecular weight of Recombinant Rat GM-CSF is Mr 14.7 kDa.
Predicted Molecular Mass
14.7
Formulation
This recombinant protein solution was 0.2 µm filtered and formulated in modified Dulbecco’s phosphate buffered saline (1X PBS) pH 7.2 – 7.3 with no calcium, magnesium, or preservatives present.
Storage and Stability
Working aliquots of this recombinant protein solution are stable for up to twelve months at -20°C to -70°C in a manual defrost freezer. Upon thawing, in the presence of a carrier protein, this recombinant protein can be stored at 2° - 8°C for one month without detectable loss of activity. For long-term storage, aliquot and freeze at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles.
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Recombinant Rat GM-CSF is widely used in research because it is a potent cytokine that regulates the growth, differentiation, and activation of multiple hematopoietic and immune cell types, making it essential for studies involving rat immune function, inflammation, hematopoiesis, and disease models.
Key scientific reasons to use recombinant rat GM-CSF in research applications include:
Stimulation of Hematopoietic Progenitors: GM-CSF promotes the proliferation and differentiation of bone marrow progenitor cells into granulocytes, monocytes/macrophages, and other myeloid lineages, which is critical for in vitro and in vivo studies of hematopoiesis and immune cell development.
Immune Cell Activation and Modulation: It acts as a survival and activation factor for mature granulocytes, monocytes/macrophages, and eosinophils, enhancing their effector functions such as phagocytosis, cytokine secretion, and antigen presentation.
Modeling Inflammatory and Infectious Diseases: GM-CSF is central to the regulation of inflammatory responses, making it valuable for modeling autoimmune diseases, allergic inflammation, and infection in rats. It has been shown to enhance host defense mechanisms and improve outcomes in infection models, such as fungal and bacterial challenges.
Therapeutic and Translational Research: Recombinant GM-CSF is used to study potential therapies for conditions involving GM-CSF deficiency or dysfunction, including immune deficiencies, chemotherapy-induced neutropenia, and as an adjuvant in cancer immunotherapy research.
Neuroimmunology and Tissue Repair: GM-CSF has roles in the central nervous system, influencing food intake, body weight, and neuroinflammation, and is involved in tissue repair and regeneration studies.
Species-Specific Activity: Using recombinant rat GM-CSF ensures optimal activity and receptor compatibility in rat models, which is crucial for accurate and reproducible results in preclinical studies.
Typical applications include:
In vitro differentiation and activation of rat bone marrow-derived macrophages or dendritic cells.
In vivo modulation of immune responses in rat models of infection, inflammation, autoimmunity, or cancer.
Functional assays to study phagocytosis, cytokine production, and immune cell signaling pathways.
In summary, recombinant rat GM-CSF is a versatile tool for dissecting immune mechanisms, modeling diseases, and evaluating therapeutic strategies in rat systems, due to its broad and potent effects on myeloid and immune cell biology.
Yes, recombinant rat GM-CSF can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to your assay system.
Supporting details:
Recombinant rat GM-CSF is widely used as an ELISA standard: Multiple technical datasheets and protocols explicitly state that recombinant rat GM-CSF is suitable as a quantitative standard for sandwich ELISA assays measuring rat GM-CSF protein levels. This is standard practice in commercial ELISA kits and custom assay development.
Assay validation and parallelism: Validation studies show that dose-response curves generated with recombinant rat GM-CSF standards are parallel to those obtained with natural rat GM-CSF, indicating that the recombinant protein is immunologically equivalent for quantification purposes in these assays. This parallelism is critical for accurate quantification of endogenous samples.
Formulation considerations: Recombinant GM-CSF is often supplied lyophilized and may include carrier proteins such as BSA for stability. For ELISA calibration, it is generally recommended to use the formulation with BSA unless your assay is sensitive to carrier proteins. Always reconstitute and dilute the standard according to the assay protocol to ensure consistency.
Assay specificity: Commercial ELISA kits for rat GM-CSF are designed to detect both natural and recombinant forms of the protein, and their antibodies are validated for this purpose.
Best practices:
Use the same buffer and diluent recommended by your ELISA protocol for reconstituting and diluting the recombinant standard.
Generate a fresh standard curve for each assay run.
Confirm that the recombinant standard matches the sequence and post-translational modifications (e.g., glycosylation) relevant to your assay, especially if you are comparing to native samples.
Limitations:
If your recombinant GM-CSF differs significantly in structure (e.g., glycosylation state) from the native protein in your samples, minor quantification discrepancies may occur. However, for most research applications, these differences are negligible when using validated commercial reagents.
Always ensure the recombinant standard is of high purity and endotoxin-free for reliable results.
Summary: Recombinant rat GM-CSF is a standard, validated choice for ELISA calibration and quantification, provided you follow assay-specific protocols and validation steps.
Recombinant Rat GM-CSF has been validated for a range of applications in published research, primarily involving functional and bioactivity assays, immunological studies, and in vivo experimentation.
Key validated applications include:
Functional/Bioassay: Used to stimulate proliferation and differentiation of hematopoietic progenitors, and to assess activation and survival of granulocytes, monocytes/macrophages, and eosinophils in vitro. This includes colony formation assays for granulocyte-macrophage progenitors and functional studies on immune cell activation.
ELISA (Enzyme-Linked Immunosorbent Assay): Employed as a quantitative standard for measuring rat GM-CSF protein levels in biological samples.
Western Blot: Used as a positive control or standard for detecting GM-CSF protein in cell lysates or tissue extracts.
Immunohistochemistry: Applied to detect GM-CSF or GM-CSF-responsive cells in tissue sections.
Blocking Assay: Utilized to block GM-CSF activity in functional studies to elucidate its biological role.
Immunoprecipitation: Used to isolate GM-CSF or GM-CSF-interacting proteins from complex mixtures.
In vivo applications in published research include:
Animal models of disease: Recombinant rat GM-CSF has been administered to rats to study its effects on immune modulation, neuroprotection, and inflammation. For example:
Modulation of microglial activation and neuroprotection in models of neurodegenerative diseases such as Parkinson’s and Alzheimer’s disease.
Regulation of food intake and body weight via central nervous system administration.
Investigation of optic nerve inflammation and demyelination in models of ischemic optic neuropathy.
Induction of M2-like macrophages for tissue repair in vivo.
Cellular assays: Used to stimulate rat immune cells (e.g., macrophages, dendritic cells, microglia) in vitro to study cytokine secretion, phagocytosis, and cell signaling pathways.
Recombinant rat GM-CSF is widely used to study hematopoiesis, immune cell function, and disease mechanisms in rat models.
It is also used as a tool to generate or manipulate specific immune cell populations (e.g., dendritic cells, macrophages) for downstream applications.
Published studies have validated its use in both in vitro and in vivo settings, supporting its reliability for translational and mechanistic research.
If you require details on a specific application or protocol, please specify the context or experimental system of interest.
To reconstitute and prepare Recombinant Rat GM-CSF protein for cell culture experiments, first briefly centrifuge the vial to collect the lyophilized powder at the bottom. Reconstitute the protein at a concentration of 50 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin (BSA) as a carrier protein to stabilize the cytokine and prevent adsorption to plastic surfaces.
Step-by-step protocol:
Centrifuge the vial before opening to ensure all lyophilized material is at the bottom.
Add sterile PBS (phosphate-buffered saline) containing at least 0.1% BSA to achieve a final concentration of 50 μg/mL.
Gently pipette the solution up and down or swirl to fully dissolve the protein; avoid vigorous vortexing to prevent denaturation.
Aliquot the reconstituted protein into small volumes to avoid repeated freeze-thaw cycles, which can degrade the protein.
Store aliquots at –20°C or colder for long-term storage; avoid repeated freeze-thaw cycles.
Additional notes:
If a different working concentration is required for your assay, further dilute the stock solution in cell culture medium or PBS with 0.1% BSA immediately before use.
For cell culture applications, ensure all solutions are sterile and endotoxin-free.
The presence of a carrier protein (BSA or HSA) is critical for stability, especially at low concentrations.
The reconstituted protein is typically stable for at least one week at 4°C and for several months at –20°C or below when aliquoted with carrier protein.
Summary Table:
Step
Details
Centrifuge vial
Collect lyophilized powder at bottom
Reconstitution buffer
Sterile PBS + ≥0.1% BSA (or HSA)
Stock concentration
50 μg/mL
Mixing
Gentle pipetting or swirling
Aliquoting
Small volumes to avoid freeze-thaw
Storage
–20°C or colder (long-term); 4°C (short-term, ≤1 week)
Working dilution
Dilute in cell culture medium or PBS + 0.1% BSA as needed
This protocol ensures optimal stability and biological activity of recombinant rat GM-CSF for cell culture experiments.
References & Citations
1. Parker, MW. et al. (2008) Cell134: 496
2. Trapnell, BC. and Whitsett, JA. (2002) Annual Review of Physiology64: 775
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
