Recombinant Mouse SCF

Recombinant Mouse SCF (Stem Cell Factor) is widely used in research because it is a critical cytokine for the survival, proliferation, and differentiation of hematopoietic stem and progenitor cells, as well as for the maintenance and expansion of other cell types in vitro.

Key scientific reasons to use recombinant Mouse SCF in your research applications:

• Hematopoietic Cell Culture: SCF is essential for the in vitro culture and expansion of hematopoietic progenitor cells, supporting both short-term proliferation and, in combination with other factors, long-term maintenance.
• Colony Formation Assays: It stimulates the growth of colony-forming cells (CFCs) from highly enriched populations, making it indispensable for colony formation and differentiation assays.
• Synergy with Other Cytokines: SCF acts synergistically with other growth factors (e.g., IL-3, GM-CSF, G-CSF) to enhance the proliferation and differentiation of myeloid, erythroid, and lymphoid progenitors.
• Mast Cell Biology: SCF is a potent mast cell chemoattractant and is required for mast cell differentiation and maintenance in vitro.
• Stem Cell Niche Regulation: SCF regulates hematopoietic stem cells (HSCs) in the bone marrow niche, contributing to their self-renewal and survival.
• Functional Assays and Receptor Signaling: It is used to investigate c-Kit (CD117) receptor signaling, which is central to many developmental and disease processes.
• Embryonic Development and Differentiation: SCF plays a pivotal role in embryonic hematopoiesis and is used to differentiate ES-derived cells toward the hematopoietic lineage.
• Additional Biological Roles: SCF is involved in melanogenesis, gametogenesis, vasculogenesis, and brain angiogenesis, making it relevant for studies beyond hematopoiesis.

Technical advantages of recombinant Mouse SCF:

• Defined Activity and Purity: Recombinant forms offer high purity (>95–97%) and well-defined biological activity, enabling reproducible and standardized experimental conditions.
• Sterility and Endotoxin Control: Research-grade preparations are sterile-filtered and have low endotoxin levels, suitable for sensitive cell culture applications.
• Soluble Form: Recombinant Mouse SCF typically corresponds to the secreted, soluble form, which is effective for short-term proliferation and functional assays.

Typical applications include:

• Expansion and maintenance of hematopoietic stem/progenitor cells
• Mast cell differentiation and migration studies
• Colony formation and differentiation assays
• Investigation of c-Kit signaling pathways
• Embryonic stem cell differentiation toward hematopoietic lineages

In summary, recombinant Mouse SCF is a foundational reagent for hematopoietic research, stem cell biology, and functional cell assays, providing reliable, standardized stimulation of c-Kit–dependent cellular processes.

Yes, recombinant mouse SCF can generally be used as a standard for quantification or calibration in ELISA assays, provided it is of high purity, its concentration is accurately known, and it matches the isoform and sequence recognized by your assay antibodies. This is a common practice in quantitative ELISA protocols for cytokines and growth factors.

Key considerations and supporting details:

• Recombinant SCF as Standard: Many commercial ELISA kits for mouse SCF use recombinant mouse SCF as the standard for generating the calibration curve. The standard is typically an affinity-purified, well-characterized recombinant protein, often corresponding to the soluble form (amino acids Lys26-Ala189 or similar).

• Purity and Formulation: The recombinant SCF used as a standard should be highly pure (typically >95–98% by SDS-PAGE) and its concentration should be accurately determined. Carrier proteins (e.g., BSA) may be included to stabilize the standard, but carrier-free formulations are preferred if BSA or other additives could interfere with your assay.

• Isoform and Sequence: Ensure the recombinant SCF matches the isoform and sequence recognized by the capture and detection antibodies in your ELISA. Most kits and protocols use the soluble form of SCF, which is the biologically active, secreted isoform.

• Validation: It is important to confirm that the recombinant SCF standard produces a standard curve that is parallel to the response obtained with endogenous SCF in your samples. This ensures accurate quantification. Some kits specifically state that their standards (recombinant SCF) yield parallel curves with natural SCF, validating their use for quantification.

• Best Practices:

  • Prepare a fresh standard curve for each assay run to ensure accuracy.
  • Use the same buffer matrix for standards and samples to minimize matrix effects.
  • Store and handle the recombinant standard according to manufacturer or protocol recommendations to maintain stability and activity.

• Limitations: Some ELISA kits may recommend against using standards from other sources or recombinant proteins not validated for their specific assay, due to potential differences in folding, glycosylation, or sequence. Always check your assay's documentation for such restrictions.

Summary Table: Recombinant Mouse SCF as ELISA Standard

In summary: If your recombinant mouse SCF meets these criteria, it is suitable for use as a standard in ELISA quantification and calibration. Always validate the standard curve and consult your assay protocol for any specific requirements or limitations.

Recombinant Mouse SCF (Stem Cell Factor) has been validated in published research for a wide range of applications, primarily related to hematopoietic stem and progenitor cell biology, cell signaling, and functional assays.

Key validated applications include:

• In vitro culture and expansion of hematopoietic progenitor and stem cells: SCF is routinely used to support the survival, proliferation, and expansion of murine hematopoietic stem cells (HSCs) and progenitors in serum-free or defined media, often in combination with other cytokines such as thrombopoietin (TPO) and FLT3 ligand.
• Colony formation assays: SCF is a critical component in methylcellulose-based colony-forming unit (CFU) assays to assess the clonogenic potential of hematopoietic progenitors.
• Differentiation of embryonic stem (ES) or induced pluripotent stem (iPS) cells: SCF is used to direct the differentiation of pluripotent stem cells toward hematopoietic lineages.
• Mast cell differentiation and maintenance: SCF is essential for the in vitro generation and maintenance of mast cells from bone marrow or progenitor sources.
• Functional bioassays: SCF is validated in cell proliferation assays, such as stimulation of TF-1 cell line proliferation, to confirm its biological activity.
• Investigation of receptor (KIT) signaling: SCF is used to study downstream signaling pathways activated by the KIT receptor in various cell types, including hematopoietic, mast, and leukemia cells.
• In vivo studies: SCF has been used in animal models to stimulate hematopoiesis, support engraftment after transplantation, and investigate its role in tissue homeostasis and disease models.
• ELISA and Western blot controls: Recombinant SCF is used as a standard or positive control in immunoassays.
• Multiplex standards and cell culture media supplementation: SCF is included in defined media for stem cell and progenitor cell research.

Representative published research applications:

• Ex vivo expansion of functional murine HSCs for transplantation and gene editing studies.
• Bioassays for cell proliferation and differentiation, including studies on leukemia, stem cell fate, and hematopoietic regulation.
• Cell culture supplementation for maintenance and expansion of hematopoietic and mast cell populations.
• Differentiation protocols for ES/iPS cells toward hematopoietic or germ cell lineages.

Additional validated uses:

• Blocking assays, immunohistochemistry, and immunoprecipitation (less common, but reported in some protocols).
• Mass spectrometry and HPLC for protein characterization and quality control.

Summary Table of Validated Applications

These applications are supported by both product validation data and peer-reviewed publications, reflecting the central role of SCF in hematopoietic and stem cell research.

To reconstitute and prepare Recombinant Mouse SCF (Stem Cell Factor) protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
2. Reconstitution:
   • For carrier-free formulations: Add sterile deionized water or sterile PBS to achieve a final concentration of 0.1–1.0 mg/mL (100–1000 μg/mL). A commonly used concentration is 100 μg/mL.
   • For formulations with carrier protein (e.g., BSA): Reconstitute in sterile PBS containing at least 0.1% BSA or serum albumin to enhance stability, especially for long-term storage or repeated freeze-thaw cycles.
   • Do not vortex; gently pipette to dissolve the protein completely.
   • Allow several minutes for complete dissolution.
3. Aliquoting:
   • Divide the reconstituted solution into working aliquots to avoid repeated freeze-thaw cycles.
   • Use low-protein-binding tubes if possible.
4. Storage:
   • Store lyophilized protein at 2–8°C or −20°C (preferably desiccated).
   • Store reconstituted aliquots at ≤−20°C (not in frost-free freezers) or −80°C for long-term storage.
   • Avoid multiple freeze-thaw cycles to preserve activity.
5. Working solution preparation:
   • For cell culture, dilute the stock solution to the desired working concentration (often in the range of 2.5–10 ng/mL for bioassays, but optimize for your specific application).
   • Make final dilutions in cell culture medium containing serum or 0.1% BSA to minimize adsorption and maintain protein stability.
6. Sterility:
   • Ensure all solutions and diluents are sterile and endotoxin-free, especially for sensitive cell types.

Summary Table: Key Steps for Recombinant Mouse SCF Reconstitution

Notes:

• Always consult the specific product datasheet for formulation details, as some preparations may require PBS instead of water, or may already contain stabilizers.
• The optimal working concentration for your cell type or assay should be determined empirically via a dose-response experiment.

These guidelines ensure maximum activity and stability of recombinant Mouse SCF for cell culture applications.