Recombinant Human Noggin (NS0 Cell Expressed)

Recombinant Human Noggin is widely used in research because it is a potent and specific antagonist of bone morphogenetic proteins (BMPs), making it essential for studies involving embryonic development, stem cell maintenance, and directed differentiation protocols.

Key reasons to use Recombinant Human Noggin in research applications:

• BMP Antagonism: Noggin binds BMPs (such as BMP-2, BMP-4, BMP-7) with high affinity, preventing them from activating their receptors. This inhibition is critical for controlling cell fate decisions, especially in neural and mesodermal differentiation.
• Stem Cell Maintenance: In human embryonic stem cell (hESC) and induced pluripotent stem cell (iPSC) cultures, Noggin is used to suppress spontaneous differentiation by antagonizing endogenous BMP signaling, thereby maintaining cells in an undifferentiated, pluripotent state.
• Directed Differentiation: Noggin is used to guide stem cells toward specific lineages. For example, it promotes neural induction by inhibiting BMPs, which otherwise drive non-neural fates. It is also used in protocols for generating retinal, endodermal, and mesodermal derivatives from pluripotent stem cells.
• Organoid and Tissue Engineering: Noggin is a key supplement in organoid cultures (e.g., brain, intestine, lung), where precise modulation of BMP signaling is required for proper tissue patterning and maturation.
• Developmental Biology: Noggin is essential for studying processes such as neural tube closure, somite formation, and limb development, as it regulates the balance between proliferation and differentiation during embryogenesis.
• Bone and Cartilage Research: Recent studies indicate Noggin can modulate osteogenic differentiation in mesenchymal stem cells and may have applications in bone regenerative therapies by influencing FGFR/Src/Akt/ERK signaling pathways.

Additional considerations:

• Functional Assays: Recombinant Noggin is used in bioassays to study BMP signaling pathways, test BMP antagonism, and investigate mechanisms of tissue homeostasis and disease.
• Quality and Consistency: Recombinant production ensures batch-to-batch consistency, purity, and defined activity, which are critical for reproducible experimental outcomes.

In summary, use Recombinant Human Noggin when you need precise, reproducible inhibition of BMP signaling to control stem cell fate, model development, or study tissue-specific differentiation and regeneration.

Yes, you can use Recombinant Human Noggin as a standard for quantification or calibration in your ELISA assays, provided it is properly purified and its concentration is accurately determined.

Key Points:

• Suitability as Standard: Recombinant Human Noggin is commonly used as a standard in ELISA assays for quantifying Noggin in biological samples. Many commercial ELISA kits recommend or include recombinant Noggin for this purpose [3, 5, 17].
• Formulation: For ELISA standard use, it is generally advised to use recombinant Noggin formulated with BSA (bovine serum albumin), as BSA helps stabilize the protein and prevents adsorption losses during dilution and assay setup [3, 5, 17].
• Carrier-Free vs. BSA-Containing: Carrier-free recombinant Noggin is recommended only if BSA could interfere with your assay (e.g., in certain cell-based assays or specific detection systems) [3, 5, 17].
• Concentration Determination: The concentration of the recombinant protein should be measured accurately, preferably by methods such as amino acid analysis or HPLC, to ensure reliable standard curve generation .
• Standard Curve Preparation: Always prepare a fresh standard curve for each ELISA run using serial dilutions of the recombinant Noggin in the same matrix as your samples (e.g., assay buffer or sample diluent) to account for matrix effects [4, 18].

Recommendations:

• Use purified recombinant Human Noggin (preferably with BSA) as the standard.
• Confirm the protein concentration before use.
• Follow the manufacturer’s instructions for reconstitution and dilution.
• Prepare a standard curve for each assay to ensure accurate quantification.

This approach will allow you to reliably quantify Noggin levels in your samples using ELISA.

Research Applications of Recombinant Human Noggin

Recombinant human Noggin has been validated across a diverse range of research applications in published studies, reflecting its multifaceted biological roles.

Stem Cell and Differentiation Studies

Noggin has been extensively utilized in stem cell research, particularly for osteogenic differentiation. Studies demonstrate that Noggin induces osteogenic gene expression in human adipose-derived stem cells, bone marrow stem cells, and dental pulp stem cells, with effective induction occurring at concentrations of 100 ng/mL. The protein promotes increased expression of osteogenic markers including RUNX2, osterix, osteonectin, bone sialoprotein, osteocalcin, and osteopontin. Additionally, Noggin supports the maintenance of undifferentiated human embryonic stem cells in vitro, preventing spontaneous differentiation when applied short-term.

Developmental and Tissue Engineering Applications

Noggin plays critical roles in developmental processes and regenerative medicine. It has been validated for use in neural tube development and patterning, as well as joint formation. In regenerative medicine contexts, Noggin has been utilized to generate cells for intestinal tissue and organoid development in vitro. Research has employed Noggin in establishing human pluripotent stem cell-derived cortical neurosphere models, small intestine-on-chip systems, and ventral midbrain-striatum-cortex assembloids.

Neuroprotection and Injury Response

Noggin demonstrates neuroprotective effects in early stages of spinal cord injury. Studies have shown that Noggin administration suppresses fibrinogen leakage into the brain during the acute phase following traumatic brain injury in animal models.

Cancer and BMP Signaling Studies

Noggin has been validated for investigating BMP signaling inhibition in cancer contexts. The protein can limit BMP signaling to help contain or reduce metastatic lesions, positioning it as a potential therapeutic research target. Additionally, Noggin can specifically activate FGFR2 in osteosarcoma cells, revealing alternative signaling mechanisms.

Functional and Bioactivity Assays

Recombinant human Noggin has been validated for bioactivity assays and neutralization studies. Functional validation includes demonstrating that Noggin inhibits alkaline phosphatase production induced by recombinant BMP-4 in chondrogenic cell lines, confirming its antagonistic relationship with BMP signaling.

To reconstitute and prepare Recombinant Human Noggin protein for cell culture experiments, follow these steps:

1. Centrifuge the vial briefly (10–30 seconds in a microcentrifuge) before opening to ensure all lyophilized protein is at the bottom.
2. Reconstitution buffer: Use sterile 1× PBS (pH 7.4) containing 0.1% human or bovine serum albumin (HSA/BSA) as a carrier protein for optimal stability and activity. Some protocols also allow reconstitution in sterile distilled water or 10 mM acetic acid, but PBS with carrier protein is preferred for cell culture applications.
3. Concentration: Reconstitute to a final concentration of 0.1–1.0 mg/mL (commonly 0.2 mg/mL or 250 μg/mL). Do not reconstitute below 100 μg/mL for stability.
4. Mixing: Gently swirl or tap the vial to dissolve the protein. Avoid vortexing or vigorous pipetting, which may denature the protein.
5. Aliquoting: Divide the reconstituted solution into small aliquots to avoid repeated freeze-thaw cycles.
6. Storage:
   • Short-term: Store at 2–8°C for up to 1 week.
   • Long-term: Store at –20°C to –80°C for up to 6 months.
   • Avoid repeated freeze-thaw cycles to preserve activity.
7. Working dilutions: Prepare further dilutions in cell culture medium or PBS containing 0.1% HSA/BSA immediately before use. The optimal working concentration should be determined empirically for your specific cell type and assay.

Summary Table:

Additional Notes:

• Always consult the specific Certificate of Analysis (CoA) for your batch for any manufacturer-specific recommendations.
• Confirm protein integrity by SDS-PAGE if needed.
• For sensitive cell types, ensure all reagents are endotoxin-free.

This protocol ensures maximum stability and biological activity of recombinant human Noggin for cell culture applications.