Members in the TNF superfamily regulate immune responses and induce apoptosis. Two novel members in the TNF family were recently identified and designated BAFF/BLyS/TALL-1/THANK/zTNF41-4 and April/TALL-2, respectively. BAFF was characterized as a B cell activator since it induced B cell proliferation and immunoglobulin secretion. April regulates immunological and non-immunological cell growth. Three receptors, BCMA (for B cell maturation protein), TACI, and BAFF-R, for BAFF were recently identified. BCMA, like TACI, binds BAFF and April. BAFF and its receptors induce B cell development and survival, activate NF-kB and c-jun N-terminal kinase, and are involved in B cell associated autoimmune diseases. BCMA expression has been found in immune organs and mature B cell lines. Although some expression has been observed at the cell surface, BCMA appears to be localized to the Golgi compartment. The binding of BCMA to APRIL or BAFF has been shown to stimulate IgM production in peripheral blood B cells and increase the survival of cultured B cells. This data suggests that BCMA may play an important role in B cell development, function and regulation. Mouse and human BCMA share 62% amino acid identity.
Protein Details
Purity
>95% by SDS-PAGE and analyzed by silver stain.
Endotoxin Level
<0.1 EU/µg as determined by the LAL method
Biological Activity
The biological activity of Mouse BCMA was determined by its ability to inhibit APRIL-mediated proliferation of anti-IgM stimulated mouse B cells. The expected ED<sub>50</sub> for this effect is 0.01 - 0.03 μg/ml in the presence of 100 ng/ml of human APRIL.
The predicted molecular weight of Recombinant Mouse BCMA is Mr 32 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is Mr 35-40 kDa.
Predicted Molecular Mass
32
Formulation
This recombinant protein was lyophilized from a 0.2 μm filtered solution in 20 mM MES, 50 mM NaCl, pH 6.2.
Storage and Stability
This lyophilized protein is stable for six to twelve months when stored desiccated at -20°C to -70°C. After aseptic reconstitution, this protein may be stored at 2°C to 8°C for one month or at -20°C to -70°C in a manual defrost freezer. Avoid Repeated Freeze Thaw Cycles. See Product Insert for exact lot specific storage instructions.
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Recombinant Mouse BCMA is a valuable tool in research applications focused on B cell biology, plasma cell maturation, and therapeutic development, particularly in the context of immunology and oncology. Its use enables precise investigation of BCMA-mediated signaling, ligand interactions, and therapeutic targeting in mouse models.
Key scientific applications and advantages:
Mechanistic Studies: Recombinant Mouse BCMA allows for detailed analysis of the molecular interactions between BCMA and its ligands, APRIL and BAFF, which are critical for plasma cell survival, maturation, and immunoglobulin production. This is essential for understanding B cell development and the regulation of antibody-secreting cells.
Therapeutic Target Validation: BCMA is highly expressed on plasma cells and multiple myeloma cells, making it a prime target for therapeutic interventions such as monoclonal antibodies, CAR-T cells, and antibody-drug conjugates. Recombinant BCMA is used to screen and characterize these agents, assess binding specificity, and evaluate functional blockade of ligand interactions.
Preclinical Model Development: In mouse models, recombinant BCMA is used to study disease mechanisms, track plasma cell maturation, and evaluate the efficacy of BCMA-targeted therapies. It is also essential for generating reporter systems and validating antibody or CAR constructs in vitro and in vivo.
Assay Development: Recombinant BCMA serves as a standard or capture antigen in ELISA, flow cytometry, and other immunoassays to quantify ligand binding, antibody affinity, or soluble BCMA levels in biological samples.
Comparative Immunology: The recombinant protein enables cross-species studies of BCMA function, given the sequence homology between mouse and human BCMA, facilitating translational research from mouse models to human disease.
Best practices:
Use recombinant BCMA in controlled binding assays to characterize ligand or antibody interactions.
Employ recombinant BCMA in functional cell-based assays to assess downstream signaling or cell survival.
Integrate recombinant BCMA into preclinical therapeutic evaluation pipelines for screening candidate molecules.
Summary: Using recombinant Mouse BCMA in research provides a robust platform for dissecting BCMA biology, validating therapeutic strategies, and developing sensitive assays, especially in mouse models of plasma cell disorders and immunological studies.
Yes, recombinant Mouse BCMA can be used as a standard for quantification or calibration in ELISA assays, provided it is well-characterized and matches the target analyte in your samples. Recombinant proteins are commonly used as standards in quantitative ELISA protocols to generate standard curves for accurate measurement of analyte concentrations.
Key considerations for using recombinant Mouse BCMA as an ELISA standard:
Purity and Characterization: The recombinant BCMA should be highly purified and its concentration accurately determined, ideally by methods such as HPLC or UV absorbance.
Sequence and Structure: The recombinant standard should match the endogenous BCMA in your samples, especially in terms of sequence and post-translational modifications, to ensure comparable antibody recognition.
Standard Curve Preparation: Prepare a dilution series of the recombinant BCMA in the same matrix as your samples (e.g., buffer, serum, or cell lysate) to account for potential matrix effects. The standard curve should cover the expected concentration range of BCMA in your samples.
Carrier Proteins: If stability is a concern, recombinant proteins with carrier proteins (e.g., BSA) can be used, but ensure that the carrier does not interfere with your assay.
Validation: Validate the standard curve for linearity, accuracy, and precision. Acceptable coefficient of variation (%CV) and relative error (%RE) should be within ±20% for quantitative assays.
Scientific best practices:
Use the same recombinant BCMA standard throughout your experiments for consistency.
Document the lot number, concentration, and preparation method of your standard.
If using a commercial ELISA kit, confirm that the kit’s antibodies recognize the recombinant BCMA standard equivalently to the endogenous protein.
In summary: Recombinant Mouse BCMA is suitable as an ELISA standard if it is properly characterized, matches the analyte, and is validated for your assay conditions. This approach is standard in quantitative ELISA workflows for cytokines, receptors, and other proteins.
Recombinant Mouse BCMA has been validated for several key applications in published research, primarily in the context of immunological and cancer studies, especially multiple myeloma.
Validated Applications:
ELISA (Enzyme-Linked Immunosorbent Assay):
Recombinant Mouse BCMA is frequently used as a capture antigen or standard in ELISA assays to detect anti-BCMA antibodies or to quantify BCMA levels in biological samples.
SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis):
The protein is validated for purity assessment and molecular weight determination by SDS-PAGE.
Flow Cytometry:
BCMA reporter mouse models utilize recombinant BCMA for tracking plasma cell development and maturation via flow cytometry.
Preclinical Therapeutic Evaluation:
Recombinant Mouse BCMA serves as a target antigen in mouse model studies for evaluating anti-BCMA therapies, including monoclonal antibodies, immunotoxins, and CAR-T cells.
It is used to validate the specificity and efficacy of therapeutic agents targeting BCMA in vitro (cell-based assays) and in vivo (mouse xenograft models).
Comparative Immunology Research:
The protein is employed in studies comparing immune responses and plasma cell biology across species.
Supporting Details:
Immunotherapy Development: Recombinant Mouse BCMA is central to the development and validation of anti-BCMA monoclonal antibodies, antibody-drug conjugates, bispecific T cell engagers (BiTEs), and CAR-T cell therapies targeting multiple myeloma and other plasma cell disorders.
Structural Biology: It is used in crystallography and binding studies to characterize antibody-BCMA interactions and to inform therapeutic design.
Functional Assays: Recombinant BCMA is used to assess ligand binding (APRIL, BAFF) and to study downstream signaling effects in B cell maturation and survival.
Summary Table of Validated Applications
Application
Description/Use Case
Reference(s)
ELISA
Detection/quantification of BCMA or anti-BCMA Abs
SDS-PAGE
Purity and molecular weight assessment
Flow Cytometry
Tracking plasma cell maturation
Preclinical Therapeutics
Target antigen for antibody, immunotoxin, CAR-T
Comparative Immunology
Cross-species immune response studies
Structural Biology
Crystallography, binding studies
Functional Assays
Ligand binding, signaling studies
These applications are supported by multiple peer-reviewed studies and are foundational for both basic research and translational therapeutic development involving BCMA.
To reconstitute and prepare Recombinant Mouse BCMA protein for cell culture experiments, follow these general steps based on best practices for recombinant protein handling:
Centrifuge the vial briefly before opening to ensure all lyophilized powder is at the bottom.
Reconstitution:
Use sterile PBS or sterile distilled water as the solvent, unless the product datasheet specifies otherwise.
Add solvent to achieve a final concentration between 0.1–1.0 mg/mL (e.g., for 100 µg protein, add 100 µL for 1 mg/mL or 1 mL for 0.1 mg/mL).
If recommended, include 0.1% BSA as a carrier protein to stabilize the solution, especially for low concentrations or if the protein is prone to adsorption.
Gently mix by pipetting up and down or by slow inversion. Do not vortex or shake vigorously, as this may denature the protein.
Incubate at room temperature for 10–15 minutes to ensure complete dissolution.
Aliquot the solution to avoid repeated freeze-thaw cycles, which can degrade the protein.
Storage:
Store aliquots at –20°C to –70°C in a manual defrost freezer for long-term storage.
For short-term use (up to one month), store at 2–8°C.
Avoid repeated freeze-thaw cycles.
Preparation for cell culture:
Dilute the reconstituted stock into your cell culture medium immediately before use.
Ensure the final working concentration is appropriate for your experimental design (commonly in the range of ng/mL to µg/mL, depending on the assay).
If using serum-containing medium, confirm compatibility and consider pre-testing for precipitation or loss of activity.
Additional notes:
Always consult the specific product datasheet for any unique instructions regarding buffer composition, concentration, or handling.
If endotoxin sensitivity is critical for your experiment, verify the endotoxin level (should be <1 EU/µg for most cell culture applications).
For functional assays, confirm protein activity post-reconstitution if possible.
Summary of key steps:
Centrifuge vial, reconstitute in sterile PBS or water (optionally with 0.1% BSA), gently mix, aliquot, store appropriately, and dilute into cell culture medium for use.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
