Recombinant Human GMFB

Recombinant Human GMFB (Glia Maturation Factor Beta) is used in research applications to study neural differentiation, neuroregeneration, and neuroinflammation, as well as to investigate its roles in cancer biology and liver regeneration. Its recombinant form offers high purity, batch-to-batch consistency, and reliable biological activity, making it a valuable tool for experimental reproducibility and mechanistic studies.

Key scientific reasons to use recombinant human GMFB in research include:

• Neural differentiation and regeneration: GMFB promotes the differentiation of brain cells and stimulates neural regeneration, making it useful for studies on neurodevelopment, neuroplasticity, and recovery from neural injury.
• Neuroinflammation and neurodegenerative disease models: GMFB is brain-specific and essential for nervous system homeostasis, offering a target for research into neurodegenerative diseases such as Alzheimer's and Parkinson's disease.
• Cancer research: GMFB has been shown to inhibit the proliferation of tumor cells, providing a model for studying tumor suppression mechanisms and potential therapeutic interventions.
• Liver regeneration: Studies using transgenic zebrafish models indicate that GMFB in hepatocytes confers protective benefits for liver regeneration and repair, supporting its use in hepatic injury and regeneration research.
• Evolutionary conservation: The amino acid sequence of GMFB is highly conserved across species, suggesting fundamental biological roles and enabling cross-species comparative studies.
• Experimental reproducibility: Recombinant proteins, including GMFB, provide high bioactivity, purity, and batch-to-batch consistency, which are critical for reproducible and interpretable experimental results.

Additional applications include:

• Investigating the molecular mechanisms of glial cell maturation and function.
• Developing assays for screening neuroprotective or neuroregenerative compounds.
• Exploring GMFB’s phosphorylation and post-translational modifications in cellular signaling pathways.

In summary, recombinant human GMFB is a versatile reagent for neuroscience, cancer, and regenerative medicine research, offering precise control over experimental variables and enabling mechanistic insights into glial biology and tissue repair.

You can use recombinant human GMFB as a standard for quantification or calibration in your ELISA assays, provided the protein is highly purified and its concentration is accurately determined.

For ELISA quantification:

• Purity and quantification: The recombinant GMFB should be of high purity (typically >90–95% by SDS-PAGE) and free from significant contaminants or endotoxins. Accurate concentration determination (e.g., by BCA assay or absorbance at 280 nm) is essential for preparing reliable standard curves.
• Standard curve preparation: Prepare a dilution series of the recombinant GMFB in the same buffer or diluent used for your samples to ensure matrix consistency. Follow best practices for reconstitution and dilution, as outlined in ELISA kit protocols or protein datasheets.
• Validation: Confirm that your ELISA antibodies recognize recombinant GMFB equivalently to endogenous GMFB. Most commercial GMFB ELISA kits are validated to detect both recombinant and native forms.
• Application: Recombinant proteins are commonly used as standards in ELISA for calibration and quantification of target analytes, as long as the protein is properly characterized and the assay is validated for this use.

Key technical considerations:

• Use a freshly prepared standard for each assay to avoid degradation or aggregation.
• Ensure the standard curve covers the expected concentration range of your samples (typically 0–1000 pg/mL, but may vary).
• Use appropriate curve-fitting methods (e.g., 4-PL or 5-PL) for data analysis.

In summary, recombinant human GMFB is suitable as an ELISA standard if it is pure, accurately quantified, and recognized by your assay antibodies. Always validate the equivalence of recombinant and endogenous GMFB in your specific assay context.

Recombinant Human GMFB (Glia Maturation Factor Beta) has been validated in published research for several key applications, primarily in functional assays, cell-based studies, and mechanistic investigations related to inflammation, cell proliferation, and neurobiology.

Key validated applications include:

• Functional assays in cell culture: Recombinant GMFB has been used to study its effects on cell viability, proliferation, apoptosis, and cytokine production. For example, in human hepatocyte and retinal pigment epithelial cell lines, recombinant GMFB was applied to assess its role in cell proliferation, inflammatory signaling (e.g., IL-6/STAT3 pathway), and response to injury or stress.

• Rescue and knockdown experiments: In mechanistic studies, recombinant GMFB is used to rescue phenotypes following GMFB knockdown or to overexpress GMFB in cell lines, allowing researchers to directly attribute observed cellular effects to GMFB activity.

• Blocking and modulation assays: Recombinant GMFB is validated for use in blocking assays to study its interaction with signaling pathways, such as actin cytoskeleton remodeling and inflammatory cascades.

• Protein-protein interaction studies: It is employed in assays to investigate binding partners and downstream signaling mechanisms, particularly in the context of actin dynamics and neuroinflammation.

• Immunological and neurobiological research: Recombinant GMFB is used to study its role in neuroprotection, neural differentiation, and modulation of inflammatory cytokine production in both neural and non-neural tissues.

• Disease modeling: It has been applied in models of liver regeneration, diabetic retinopathy, and neurodegenerative diseases to elucidate its functional impact and therapeutic potential.

Experimental techniques where recombinant GMFB has been validated include:

• Western blotting (to confirm protein expression and modulation)
• ELISA (to measure cytokine levels)
• Flow cytometry (to assess apoptosis)
• Cell viability assays (e.g., CCK-8)
• Immunohistochemistry (for tissue localization)
• Transfection-based overexpression and knockdown studies

Summary of research contexts:

• Liver regeneration: Recombinant GMFB was used to demonstrate its role in promoting hepatocyte proliferation and modulating inflammatory responses in mouse and human cell models.
• Retinal cell injury: It was validated in studies of high glucose-induced injury in human retinal pigment epithelial cells, showing effects on inflammation and apoptosis.
• Neurobiology: Applications include studies on neural differentiation, neuroprotection, and cytokine modulation in astrocytes and microglia.

These applications are supported by both primary research articles and technical datasheets from protein suppliers, which confirm the use of recombinant human GMFB in functional and blocking assays, as well as in a variety of cell-based and biochemical studies.

To reconstitute and prepare Recombinant Human GMFB protein for cell culture experiments, dissolve the lyophilized protein in sterile deionized water at a concentration of at least 0.1–0.5 mg/mL (100–500 µg/mL).

Step-by-step protocol:

1. Equilibrate the vial and diluent (sterile deionized water or PBS, pH 7.2–7.4) to room temperature before opening.
2. Centrifuge the vial briefly to collect all lyophilized material at the bottom before opening.
3. Add sterile deionized water (or PBS if specified by your protocol) to achieve a final concentration of 0.1–0.5 mg/mL. For some applications, concentrations up to 1 mg/mL are acceptable.
4. Gently mix by swirling or inverting the vial. Avoid vigorous pipetting or vortexing to prevent protein denaturation.
5. Allow the protein to dissolve for 15–30 minutes at room temperature. If undissolved material remains, gently mix for up to 2 hours.
6. Optional: For increased stability, especially for long-term storage or repeated freeze/thaw cycles, add a carrier protein or stabilizer such as 0.1% BSA, 5% HSA, 10% FBS, 5% trehalose, or 5–50% glycerol.
7. Aliquot the reconstituted protein to avoid repeated freeze/thaw cycles.
8. Storage:
   • Store aliquots at –20°C for up to 3 months.
   • At 2–8°C, the solution is stable for up to 1 week.
   • Avoid repeated freeze/thaw cycles.

Preparation for cell culture:

• Before adding to cells, dilute the stock solution to the desired working concentration using sterile cell culture medium or buffer compatible with your assay.
• If using additives (e.g., BSA, FBS), ensure they are compatible with your downstream application and cell type.

Notes:

• Always use sterile technique to prevent contamination.
• If the protein was lyophilized from PBS, reconstitution in water is generally sufficient, but check your specific product datasheet for buffer compatibility.
• For sensitive applications, consider filtering the reconstituted solution through a 0.22 µm sterile filter.

Summary Table:

These steps will ensure optimal solubility and stability of recombinant GMFB for cell culture experiments.