Recombinant Rat IL-3

Recombinant Rat IL-3 is used in research applications to study and manipulate hematopoiesis, immune cell function, and related signaling pathways in rat models. It is a potent cytokine that stimulates the proliferation, differentiation, and functional activation of various hematopoietic and immune cells.

Key reasons to use recombinant rat IL-3 in your research:

• Hematopoietic Stem and Progenitor Cell Studies: IL-3 promotes the proliferation and differentiation of pluripotent hematopoietic stem cells and lineage-committed progenitors, including those for neutrophils, macrophages, megakaryocytes, and erythroid cells. This makes it essential for in vitro colony-forming assays and studies of bone marrow cell development.

• Immune Cell Activation and Function: IL-3 augments the growth and function of mature macrophages, mast cells, eosinophils, basophils, and megakaryocytes. It is commonly used to generate and maintain cultures of these cell types, as well as to study their activation, degranulation, and cytokine release.

• Species-Specific Activity: Recombinant rat IL-3 is species-specific, ensuring physiological relevance when working with rat cells or in vivo rat models. This is important for translational research and for avoiding cross-species artifacts.

• Dendritic Cell Generation: In combination with other cytokines (e.g., CD40L, IL-4), IL-3 can drive the production of dendritic cells from progenitors, supporting studies in antigen presentation and immune regulation.

• Recovery from Marrow Suppression: Recombinant IL-3 has been shown to augment recovery in states of natural or treatment-related marrow failure, making it valuable for regenerative medicine and transplantation research.

• Bioassay Standardization: Recombinant proteins provide consistent, high-purity, and well-characterized reagents, which are critical for reproducibility in bioassays and mechanistic studies.

• Neuroscience and Inflammation: IL-3 also has roles in the nervous system and in the regulation of chronic inflammatory diseases, broadening its utility to neuroimmunology and inflammation research.

In summary, recombinant rat IL-3 is a versatile tool for dissecting hematopoietic and immune processes in rat systems, supporting applications from basic cell biology to disease modeling and therapeutic development.

Yes, recombinant rat IL-3 can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system. Most commercial rat IL-3 ELISA kits are designed to quantify both natural and recombinant rat IL-3, and they typically use recombinant IL-3 as the standard for generating the calibration curve.

Key considerations:

• Formulation: Recombinant rat IL-3 is available in both carrier-free and carrier-added (e.g., with BSA) forms. For use as an ELISA standard, the formulation with BSA is often recommended because it enhances protein stability and mimics the matrix of biological samples, reducing adsorption to plasticware and improving reproducibility.
• Reconstitution: Follow the manufacturer’s instructions for reconstitution, typically in PBS with at least 0.1% BSA or another suitable protein carrier to maintain stability and prevent loss due to adsorption.
• Assay Compatibility: Ensure that the recombinant IL-3 standard is recognized by the antibodies used in your ELISA. Most commercial kits specify that their antibodies detect both natural and recombinant forms, but if you are developing your own assay, confirm cross-reactivity and parallelism between the standard and your samples.
• Standard Curve Preparation: Prepare a serial dilution of the recombinant IL-3 standard according to your ELISA protocol to generate a standard curve for quantification.
• Storage: Store the reconstituted standard at -20°C or -80°C as recommended to maintain stability.

Limitations:

• The recombinant standard should be from the same species (rat IL-3 for rat samples) and ideally the same isoform as the endogenous protein in your samples to ensure accurate quantification.
• If using a carrier-free standard, take extra care to prevent protein loss due to adsorption, especially at low concentrations.

Summary Table: Use of Recombinant Rat IL-3 as ELISA Standard

In summary: Recombinant rat IL-3 is widely used as a standard in ELISA assays for quantification and calibration, provided it matches the assay requirements and is handled according to best practices.

Recombinant Rat IL-3 has been validated in published research primarily for applications involving the stimulation, proliferation, and differentiation of hematopoietic cells, as well as for use in cell-based bioassays.

Key validated applications include:

• Cellular Bioassays: Recombinant rat IL-3 has been used to stimulate proliferation and differentiation of rat bone marrow-derived mast cells, as demonstrated in studies assessing mast cell degranulation and cytokine release. This application is supported by published research, such as the study by Huang et al. (2014), which used recombinant rat IL-3 in bioassays to investigate TRPM7 channel function in mast cells.

• Hematopoietic Cell Culture: IL-3 is widely used to support the growth, survival, and differentiation of pluripotent hematopoietic stem cells and lineage-committed progenitors, including neutrophils, macrophages, megakaryocytes, and erythroid cells. It is also used to augment the functional activity of basophils, mast cells, eosinophils, and macrophages in vitro.

• Generation of Dendritic Cells: In combination with other cytokines (such as CD40L or IL-4), recombinant rat IL-3 has been used to stimulate the production of dendritic cells from progenitor populations.

• ELISA Standard: The protein is recommended as a standard in ELISA assays for quantifying IL-3 or for validating antibody-based detection systems.

• Functional Assays: Recombinant rat IL-3 is validated for use in functional assays to assess its biological activity, such as measuring the proliferation of responsive cell lines or primary cells.

Summary Table: Validated Applications for Recombinant Rat IL-3

Additional Notes:

• Recombinant rat IL-3 is generally considered species-specific in its activity, and its use is validated primarily in rat cell systems.
• The protein is available in both carrier-free and BSA-containing formulations, with the carrier-free version recommended when BSA could interfere with downstream applications.
• Published research supports its use in both basic research and preclinical studies involving rat hematopoietic and immune cell biology.

If you require details on a specific experimental protocol or cell type, please specify for more targeted information.

To reconstitute and prepare Recombinant Rat IL-3 protein for cell culture experiments, first determine whether your protein is carrier-free or contains a carrier protein (such as BSA), as this affects the choice of reconstitution buffer.

General Protocol for Carrier-Free Recombinant Rat IL-3:

• Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
• Reconstitute the protein in sterile distilled water or sterile PBS to a concentration of 100 μg/mL (0.1 mg/mL) or as recommended by the manufacturer.
• Gently pipet to dissolve the protein, washing down the sides of the vial to maximize recovery.
• If required for stability, further dilute in cell culture medium or PBS containing 0.1% BSA or another carrier protein.
• Aliquot the solution to avoid repeated freeze-thaw cycles.
• Store aliquots at –20°C to –70°C in a manual defrost freezer. Avoid frost-free freezers and repeated freeze-thaw cycles.

General Protocol for Recombinant Rat IL-3 with Carrier Protein (e.g., BSA):

• Reconstitute in sterile PBS containing at least 0.1% human or bovine serum albumin to a concentration of 100 μg/mL.
• Aliquot and store as above.

Preparation for Cell Culture:

• Before use, dilute the reconstituted stock into your cell culture medium to the desired working concentration (typically in the range of 0.1–10 ng/mL, but optimal concentration should be determined empirically for your assay).
• Always use sterile technique throughout the process to prevent contamination.

Additional Best Practices:

• If the protein is sensitive, minimize exposure to room temperature and avoid vigorous mixing.
• For dose-response experiments, prepare serial dilutions in medium containing carrier protein to maintain stability and minimize adsorption to plasticware.
• If endotoxin sensitivity is a concern, confirm the endotoxin level is suitable for your application (typically <0.1 ng/μg for cell culture).

Summary Table:

Key Steps:

• Centrifuge vial before opening.
• Reconstitute at 100 μg/mL in appropriate buffer.
• Aliquot and store at –20°C to –70°C.
• Dilute into cell culture medium for experiments.

Always consult the specific product datasheet for any manufacturer-specific recommendations regarding buffer composition, concentration, and storage.