Recombinant Human TSG-6

Recombinant Human TSG-6 is widely used in research applications due to its potent anti-inflammatory, tissue-protective, and immunomodulatory properties, making it a valuable tool for studying inflammation, tissue repair, and extracellular matrix biology.

Key scientific reasons to use recombinant TSG-6 in research:

• Anti-inflammatory effects: TSG-6 suppresses inflammation by modulating immune cell activity, inhibiting neutrophil and macrophage infiltration, and regulating chemokine function. It is a key mediator of the therapeutic effects of mesenchymal stem cells (MSCs) in models of myocardial infarction, corneal injury, peritonitis, lung injury, and autoimmune diseases.

• Tissue protection and repair: Recombinant TSG-6 promotes wound healing, reduces tissue damage, and inhibits fibrosis in various models, including ocular, renal, and joint tissues. It can decrease matrix-degrading enzyme production and cartilage breakdown, making it relevant for osteoarthritis and other degenerative conditions.

• Mechanistic studies: Using recombinant TSG-6 allows precise dissection of its molecular mechanisms, such as its interaction with hyaluronan and CD44, modulation of TLR/NF-κB signaling, and downstream effects on inflammatory pathways.

• Biomarker and therapeutic candidate: TSG-6 serves as a biomarker for MSC efficacy and disease progression (e.g., osteoarthritis), and recombinant protein administration can reproduce many beneficial effects of MSC therapy, enabling controlled experimental interventions.

• Versatile applications: Recombinant TSG-6 is suitable for a range of experimental protocols, including cell culture, bioassays, ELISA standards, in vivo models, and extracellular matrix studies.

In summary, recombinant human TSG-6 is a scientifically validated reagent for investigating inflammation, tissue repair, and extracellular matrix biology, and for developing novel therapeutic strategies targeting these processes.

Yes, you can use recombinant human TSG-6 as a standard for quantification or calibration in your ELISA assays, provided it is of high purity and its concentration is accurately known. Recombinant proteins are commonly used as standards in quantitative ELISA assays to generate standard curves, which are essential for determining the concentration of TSG-6 in your samples.

Key considerations and best practices:

• Purity and Formulation: The recombinant TSG-6 should be highly purified and, ideally, carrier-free if you want to avoid interference from stabilizers like BSA. Some suppliers offer both carrier-free and BSA-containing formulations; the latter is often recommended for use as an ELISA standard to improve stability.
• Concentration Verification: Ensure the protein concentration is accurately determined, typically by absorbance at 280 nm or another validated method.
• Standard Curve Preparation: Prepare a serial dilution of the recombinant TSG-6 in the same buffer as your samples to generate a standard curve. This allows for precise quantification of unknowns by comparison to the standard curve.
• Compatibility: Confirm that the recombinant TSG-6 is recognized by the antibodies used in your ELISA. Most commercial ELISA kits for human TSG-6 are validated to detect both natural and recombinant forms of the protein.
• Range and Sensitivity: The standard curve should cover the expected concentration range of TSG-6 in your samples. For TSG-6 ELISAs, typical standard curve ranges are from 0.2 ng/mL to 50 ng/mL or higher, depending on the kit and assay sensitivity.

Scientific context:

• Recombinant human TSG-6 has been used as a standard in published ELISA protocols for quantifying TSG-6 in cell culture supernatants, plasma, and serum.
• The use of recombinant standards is a well-established practice in quantitative immunoassays, as long as the recombinant protein is structurally and immunologically equivalent to the native protein.

Summary of best practices:

• Use a highly purified, well-characterized recombinant human TSG-6.
• Prepare the standard curve in the same matrix as your samples.
• Validate that your ELISA antibodies detect the recombinant standard equivalently to native TSG-6.
• Follow the ELISA kit or assay protocol for standard preparation and curve fitting.

If you follow these guidelines, recombinant human TSG-6 is suitable and widely accepted as a standard for ELISA quantification of TSG-6.

Research Applications of Recombinant Human TSG-6

Recombinant human TSG-6 (tumor necrosis factor-α-stimulated gene/protein-6, also known as TNFAIP6) has been validated across diverse therapeutic and research applications in published studies.

Anti-inflammatory and Immunomodulatory Applications

The primary validated application of recombinant TSG-6 is suppression of sterile inflammation. In multiple in vivo models, recombinant TSG-6 has demonstrated efficacy in reducing inflammatory responses, including in corneal chemical injury, sterile peritonitis, and bleomycin-induced lung injury. The protein functions as a key mediator of mesenchymal stem cell (MSC) therapeutic effects across various autoimmune diseases.

Specifically, recombinant TSG-6 has been shown to attenuate severe burn-induced excessive inflammation by inhibiting activation of P38 and JNK signaling pathways. In thyroid-associated orbital inflammation models, TSG-6 treatment reduced T cell infiltration and orbital fibrosis, demonstrating superior efficacy compared to dexamethasone in inhibiting fibrotic responses.

Tissue Protection and Fibrosis Modulation

Recombinant TSG-6 protein has been validated for inhibiting fibroblast growth and fibrotic responses. The protein suppresses fibroblast proliferation through the TGF-β/Smad2 signaling pathway, with applications in conditions involving excessive fibrosis. Additionally, the link module of TSG-6 has demonstrated efficacy in promoting wound healing and suppressing inflammation in dry eye disease models.

Bioassay and Functional Assay Applications

Recombinant TSG-6 is validated for use in blocking assays and functional assays, making it suitable for mechanistic studies investigating inflammatory pathways and protein-protein interactions. The protein also serves as a potential biomarker for disease activity and progression assessment in inflammatory conditions.

Extracellular Vesicle Research

Recent applications include investigation of TSG-6-enriched extracellular vesicles as mediators of cell-to-cell communication in fibrotic disease pathophysiology, expanding the protein's utility in understanding intercellular signaling mechanisms.

To reconstitute and prepare Recombinant Human TSG-6 protein for cell culture experiments, dissolve the lyophilized protein at a concentration of 100 μg/mL in sterile PBS.

Essential steps and best practices:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (not water, unless otherwise specified in the product datasheet) to achieve the desired concentration, typically 100 μg/mL.
• Gently mix by pipetting or swirling; avoid vigorous vortexing to prevent protein denaturation.
• Aliquot the reconstituted protein to minimize freeze-thaw cycles, which can degrade protein activity.
• Storage after reconstitution: Store aliquots at 2–8 °C for up to 1 month under sterile conditions, or at –20 °C to –70 °C for longer-term storage.
• Avoid repeated freeze-thaw cycles to maintain protein integrity.

Additional recommendations for cell culture:

• If using for sensitive cell types or low-concentration applications, consider adding a carrier protein (e.g., 0.1–1% BSA or HSA) to prevent adsorption to plastic surfaces, unless this interferes with your assay.
• Filter-sterilize the final solution if sterility is a concern and the buffer or protein source is not already sterile.
• Prepare working dilutions in cell culture medium immediately before use to avoid prolonged storage at low concentrations.

Summary protocol:

1. Briefly centrifuge the vial.
2. Add sterile PBS to reach 100 μg/mL.
3. Mix gently until fully dissolved.
4. Aliquot and store as recommended.
5. Prepare working dilutions fresh for each experiment.

Always consult the specific product datasheet for any unique instructions or buffer requirements, as some recombinant proteins may have formulation-specific needs.