Recombinant Human BAFF-R

Recombinant Human BAFF-R is a valuable tool for research applications due to its central role in B-cell biology and immune regulation. Here are several reasons why you should consider using it in your studies:

1. Study of B-Cell Survival and Maturation

Recombinant Human BAFF-R allows you to investigate the mechanisms underlying B-cell survival, maturation, and antibody production. BAFF-R signaling is essential for the development and maintenance of mature B cells, making it critical for understanding normal and pathological B-cell responses.

2. Investigation of NF-κB Signaling Pathway

BAFF-R is a key activator of the NF-κB signaling pathway, which regulates gene transcription important for immune regulation and cell survival. Using recombinant BAFF-R enables you to dissect the molecular events involved in this pathway and its impact on B-cell function.

3. Modeling Autoimmune Diseases

Dysregulation of the BAFF/BAFF-R axis has been implicated in autoimmune diseases such as systemic lupus erythematosus, rheumatoid arthritis, and Sjögren’s syndrome. Recombinant BAFF-R can be used to model these conditions and explore potential therapeutic interventions targeting this pathway.

4. Research on B-Cell Malignancies

BAFF-R is expressed on most malignant B cells, including those found in primary central nervous system lymphoma (PCNSL) and diffuse large B-cell lymphoma (DLBCL). Knockout or inhibition of BAFF-R has been shown to slow tumor growth and prolong survival in preclinical models, highlighting its potential as a therapeutic target for B-cell cancers.

5. Development of Immunotherapies

Recombinant BAFF-R can be used to develop and test novel immunotherapeutic strategies, such as BAFF-based CAR-T cells, which have demonstrated efficacy in killing multiple B-cell cancer types. This makes it an important reagent for advancing cancer immunotherapy research.

6. Functional Assays and Drug Screening

Recombinant BAFF-R is suitable for various functional assays, including binding studies, signal transduction experiments, and drug screening. It can help identify modulators of BAFF-R activity, which may lead to new treatments for autoimmune diseases and B-cell malignancies.

7. Standardization and Reproducibility

Using recombinant BAFF-R ensures consistency and reproducibility in your experiments, as it provides a well-characterized and standardized source of the protein. This is particularly important for comparative studies and high-throughput screening.

8. Support for Preclinical Evaluation

Recombinant BAFF-R supports the preclinical evaluation of combinatory treatment protocols targeting the BAFF/BAFF-R axis. This is crucial for translating basic research findings into clinical applications.

In summary, recombinant Human BAFF-R is a versatile and essential reagent for studying B-cell biology, immune regulation, autoimmune diseases, and B-cell malignancies. Its use can enhance the depth and breadth of your research, providing insights that may lead to new therapeutic strategies.

Recombinant Human BAFF-R can be used as a standard for quantification or calibration in ELISA assays, provided it is highly purified, its concentration is accurately determined, and it is compatible with your assay system.

Key considerations and supporting details:

• Purity and Quantification: For ELISA standard curves, a purified protein is recommended. If a highly purified native protein is unavailable, a recombinant protein is commonly used, provided its concentration is accurately measured (e.g., by HPLC or absorbance at 280 nm). This ensures the standard curve reflects the true analyte concentration.

• Assay Compatibility: The recombinant standard must be recognized by the capture and detection antibodies used in your ELISA. Most commercial BAFF-R ELISA kits use recombinant BAFF-R as the standard, and their protocols specify that the kit is validated for both natural and recombinant forms. However, you must confirm that your recombinant BAFF-R matches the epitope(s) recognized by your assay antibodies.

• Matrix Effects: The standard should be diluted in the same buffer or matrix as your samples to minimize matrix effects and ensure accurate quantification. If your samples are in serum, plasma, or cell culture supernatant, prepare your standard curve in the same matrix or with an appropriate diluent.

• Validation: It is essential to validate the linearity and parallelism of your standard curve using the recombinant BAFF-R. This means that serial dilutions of your recombinant standard should produce a linear response, and the curve should be parallel to that obtained with endogenous BAFF-R in your sample matrix.

• Interference: Some ELISA kits report that recombinant BAFF-R can interfere with BAFF quantification at high concentrations (e.g., above 25 ng/mL), but this is specific to BAFF detection, not BAFF-R detection. For BAFF-R quantification, recombinant BAFF-R is the intended standard.

• Documentation: Always document the source, lot, and quantification method for your recombinant standard, and include this information in your assay records for reproducibility.

Summary Table: Use of Recombinant Human BAFF-R as ELISA Standard

In summary: You can use recombinant human BAFF-R as a standard for ELISA quantification if it is pure, accurately quantified, and validated for your assay system. Always perform appropriate controls and validation steps to ensure reliable quantification.

Recombinant Human BAFF-R has been validated in published research for several key applications, primarily in the context of immunology, oncology, and therapeutic antibody development.

Validated Applications in Published Research:

• Bioassays: Recombinant BAFF-R is widely used in bioassays to study ligand-receptor interactions, B-cell signaling, and proliferation. For example, it has been used to demonstrate BAFF-induced proliferation of human mesangial cells and to assess the functional activity of BAFF and its inhibitors.

• ELISA (Enzyme-Linked Immunosorbent Assay): Recombinant BAFF-R serves as a capture reagent in ELISA to quantify BAFF levels in biological samples or to screen for BAFF-R binding antibodies.

• Therapeutic Antibody Validation: Recombinant BAFF-R has been used as an immunogen or screening target for the development and validation of monoclonal antibodies (mAbs) targeting BAFF-R, which are being investigated for the treatment of B-cell malignancies and autoimmune diseases. These studies include both in vitro cytotoxicity assays and in vivo efficacy models (e.g., xenograft mouse models of lymphoma, chronic lymphocytic leukemia, and acute lymphoblastic leukemia).

• Functional Assays: It is used in functional assays to assess the biological activity of BAFF-R signaling, such as activation of the NF-κB pathway, B-cell survival, and antibody class switching.

• Blocking Assays: Recombinant BAFF-R is employed in blocking assays to evaluate the inhibitory potential of candidate molecules or antibodies that disrupt BAFF/BAFF-R interactions.

• Cellular and Molecular Characterization: Recombinant BAFF-R is used to characterize receptor expression, ligand specificity, and downstream signaling events in various cell types, including B cells and engineered cell lines.

Key Research Contexts:

• Cancer Immunotherapy: Recombinant BAFF-R has been central to the development of novel immunotherapies, such as anti-BAFF-R monoclonal antibodies and BAFF ligand-based CAR-T cells, validated in preclinical and early clinical studies for B-cell malignancies (e.g., lymphoma, multiple myeloma, acute lymphoblastic leukemia).

• Autoimmune Disease Models: It has been used in preclinical models (e.g., collagen-induced arthritis) to validate the therapeutic potential of BAFF-R-targeting antibodies for autoimmune diseases like rheumatoid arthritis.

• B-cell Biology: Recombinant BAFF-R is a tool for dissecting the role of BAFF-R in B-cell maturation, survival, and immune regulation, both in normal physiology and disease states.

Summary Table:

These applications are well-supported by published studies and product validation data, reflecting the broad utility of recombinant human BAFF-R in both basic and translational immunology research.

To reconstitute and prepare Recombinant Human BAFF-R protein for cell culture experiments, follow these best-practice steps:

1. Centrifuge the vial before opening to ensure all lyophilized protein is at the bottom.
2. Reconstitute the protein using sterile water or sterile PBS, as specified in the product datasheet or Certificate of Analysis (CoA). A typical concentration range is 0.1–1.0 mg/mL. For example, add 100 μL–1 mL of sterile water to 100 μg of protein to achieve 1 mg/mL–0.1 mg/mL.
3. Gently mix by swirling or pipetting up and down. Avoid vigorous shaking or vortexing to prevent protein denaturation or foaming.
4. Allow the protein to dissolve at room temperature for 15–30 minutes with gentle agitation.
5. Dilute for cell culture: For working solutions, further dilute the reconstituted protein in cell culture medium containing a carrier protein such as 0.1% BSA or 5–10% FBS to prevent adsorption and loss of activity, especially at low concentrations. If using serum-free conditions, use a non-animal stabilizer like trehalose.
6. Aliquot and store:
   • For short-term use (up to 1 week), store at 2–8°C.
   • For long-term storage, aliquot and freeze at –20°C to –80°C. Avoid repeated freeze-thaw cycles.
   • Optionally, add 5–50% glycerol for enhanced stability during long-term storage.

Additional notes:

• Always consult the specific product datasheet or CoA for any unique requirements, such as buffer composition or reconstitution volume.
• If the protein is supplied carrier-free and you require high stability or low working concentrations, include a carrier protein in all dilutions.
• For functional assays, typical working concentrations for BAFF-R in cell culture may range from 20–80 ng/mL in the presence of BAFF, but this should be optimized for your specific application.

Summary of key steps:

• Centrifuge vial → Add sterile water/PBS (0.1–1.0 mg/mL) → Gently dissolve → Dilute in medium with carrier protein → Aliquot and store appropriately.

These guidelines ensure maximal protein stability and activity for cell culture experiments.