Recombinant Human BAFF

Recombinant Human BAFF is used in research applications to study and manipulate B cell survival, maturation, activation, and immune regulation, making it a critical tool for immunology, oncology, and autoimmune disease research.

Key scientific reasons to use Recombinant Human BAFF:

• B Cell Biology: BAFF (B cell activating factor, also known as BLyS or TNFSF13B) is essential for the survival, maturation, and differentiation of peripheral B cells. It supports B cell proliferation and prevents apoptosis, allowing for the maintenance and expansion of B cell populations in vitro.
• Immune Modulation: BAFF not only affects B cells but also co-stimulates T cell activation, promotes a Th1-biased immune response, and expands regulatory T cells (Tregs). It also influences monocyte survival and macrophage differentiation, making it valuable for studying broader immune interactions.
• Disease Modeling: Elevated BAFF levels are associated with B cell malignancies, autoimmune diseases (such as systemic lupus erythematosus and rheumatoid arthritis), and transplant rejection. Recombinant BAFF enables the modeling of these disease states in vitro and the testing of therapeutic interventions targeting BAFF signaling.
• Antigen Presentation and Immunotherapy: BAFF-activated B cells show enhanced antigen-presenting capacity, which can be exploited to generate potent antigen-specific cytotoxic T lymphocyte (CTL) responses for cancer immunotherapy or chronic viral infection models.
• Functional Assays and Differentiation Studies: Recombinant BAFF is used in bioassays to assess B cell function, antibody production, and class switching, as well as in studies of TNF-receptor superfamily signaling.
• Standardization and Reproducibility: Recombinant proteins provide consistent, defined activity, reducing variability compared to native or serum-derived factors, which is crucial for reproducible experimental results.

Typical applications include:

• B cell culture and expansion
• Differentiation assays (e.g., plasma cell generation)
• Immune cell signaling studies
• Disease modeling (autoimmunity, lymphoma, immunodeficiency)
• Functional bioassays (e.g., proliferation, survival, cytokine secretion)
• Antigen presentation and CTL generation protocols

In summary, Recombinant Human BAFF is indispensable for dissecting B cell biology, modeling immune-related diseases, and developing immunotherapies, due to its well-characterized, potent, and reproducible effects on immune cell populations.

Yes, you can use recombinant human BAFF as a standard for quantification or calibration in your ELISA assays, provided that:

• The recombinant BAFF is of high purity and its concentration is accurately known.
• The recombinant BAFF is compatible with the specific antibodies used in your ELISA kit (i.e., it is recognized by both the capture and detection antibodies).
• The recombinant BAFF standard is prepared in a matrix similar to your samples to minimize matrix effects.

Many commercial ELISA kits for human BAFF use recombinant human BAFF as the standard (often expressed in NS0 or other systems), and these kits are calibrated against such standards. For example, the R&D Systems Quantikine ELISA Kit and several other kits referenced use NS0-expressed recombinant human BAFF as the standard.

However, always check the manufacturer’s instructions for your specific ELISA kit, as some kits may recommend or require the use of their proprietary standard for optimal accuracy and reproducibility. Using a non-kit standard may introduce variability if not properly validated.

Recombinant Human BAFF has been validated in published research primarily for bioassays, functional assays, and cell culture studies focused on B cell biology, immune regulation, and disease modeling.

Key validated applications include:

• Bioassays: Used to assess B cell activation, proliferation, differentiation, and survival in vitro. BAFF stimulates B cell lineage cells, promotes maturation, and supports peripheral B cell survival.
• Functional assays: Applied to study the signaling of TNF-receptor superfamily members (BAFF-R, TACI, BCMA) and their downstream effects on B cells and plasma cells.
• Cell culture: Utilized to maintain and differentiate B cells, investigate antibody production, and model immune responses.
• Immunoregulatory studies: BAFF has been used to explore its role in T cell co-stimulation, Th1 immune bias, Treg expansion, and monocyte/macrophage differentiation.
• Disease modeling: Validated in research on autoimmune diseases (e.g., systemic lupus erythematosus, rheumatoid arthritis), B cell malignancies, and transplant immunology.
• Antitumor immunity: BAFF has been shown to regulate monocytes in the tumor microenvironment and enhance antitumor immune responses in melanoma models.
• Antibody production: Studies have used BAFF to investigate metabolic reprogramming required for antibody production and plasma cell differentiation.
• Blocking/neutralization assays: Recombinant BAFF is used in blocking assays to study receptor-ligand interactions and in neutralization experiments to assess functional inhibition.

Additional validated uses in published research:

• ELISA: Used as a standard or analyte in immunoassays to quantify BAFF or its receptors.
• Investigation of B cell-related pathologies: BAFF is elevated in B cell malignancies, autoimmunity, and immune disorders, and recombinant BAFF is used to model these conditions in vitro.
• Synergistic studies: BAFF has been combined with other cytokines (e.g., IL-21) to study synergistic effects on plasma cell differentiation.

In summary, recombinant human BAFF is a well-validated tool for studying B cell biology, immune regulation, and related disease mechanisms in a variety of experimental formats, especially bioassays and functional cell-based assays.

To reconstitute and prepare Recombinant Human BAFF protein for cell culture experiments, follow these best practices based on manufacturer guidelines and scientific protocols:

1. Reconstitution

• Centrifuge the vial briefly (e.g., 3000–3500 rpm for 5 minutes) before opening to ensure the lyophilized powder is at the bottom.
• Reconstitute the protein with sterile PBS or the recommended buffer (often sterile PBS containing at least 0.1% human or bovine serum albumin, BSA). Typical reconstitution concentration is 100 µg/mL.
• Gently dissolve the protein by slowly adding the buffer and allowing it to sit at room temperature for 15–30 minutes. Avoid vortexing or vigorous shaking to prevent protein denaturation.
• Gently mix by pipetting up and down or tapping the vial.

2. Storage

• Short-term storage: Store the reconstituted protein at 2–8°C for up to one week.
• Long-term storage: For longer storage, dilute the protein in a buffer containing a carrier protein (e.g., 0.1% BSA, 10% FBS, or 5% HSA) and aliquot. Store aliquots at –20°C to –80°C. Avoid repeated freeze-thaw cycles.

3. Dilution for Cell Culture

• Dilute the reconstituted protein in cell culture medium or an appropriate buffer (such as PBS or medium containing carrier protein) to the desired working concentration.
• Carrier proteins (e.g., BSA, FBS, HSA) help stabilize the protein and prevent adsorption to surfaces.
• For serum-free or in vivo experiments, use trehalose as a stabilizer instead of animal-derived proteins.

4. Handling Tips

• Always use sterile technique.
• Aliquot the protein to minimize freeze-thaw cycles.
• Follow the specific instructions provided in the Certificate of Analysis (CoA) or product manual for optimal results.

These steps ensure the protein remains active and suitable for cell culture experiments.