The predicted molecular weight of Recombinant Human M2BP, MAC-2BP, Galectin-3BP is 65 kDa. However, the actual molecular weight as observed by migration on SDS-PAGE is 26-30 kDa, 60-69 kDa and 81-94 kDa (reducing conditions).
Predicted Molecular Mass
65 kDa
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS and DTT with Trehalose.
Reconstitution
Reconstitute in PBS to 500 μg/mL
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.
Applications and Recommended Usage ? (Quality Tested by Leinco)
ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 5 µg/ml (100 µl/well) µg/ml. Flow Cytometry: PN:A106 Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing CXCR5. A suggested method would be to stain cells expressing CXCR5 with approximately 10 µl per test. A typical test sample constitutes approximately 50 µl of packed whole blood or 1 x 105 continuous passage or activated cell cultures that have been centrifuged at 500 X g for five minutes. Labeling of the cells with the biotin conjugate should be followed by PN:A104, resuspended in 200-400 µl of 1X PBS.
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Recombinant Human Galectin-3BP is a multifunctional glycoprotein widely used in research due to its roles in cell–cell and cell–matrix interactions, immune modulation, cancer biology, and viral infection mechanisms. Its use enables precise mechanistic studies, biomarker validation, and therapeutic target exploration in diverse biomedical fields.
Key scientific reasons to use recombinant Human Galectin-3BP in research applications:
Modeling Cell Signaling and Immune Responses: Galectin-3BP regulates antiviral immune responses by interacting with galectin-3 and activating TNF receptor-associated factors, leading to the induction of interferons and proinflammatory cytokines such as IL-6. This makes it valuable for dissecting innate immunity and inflammatory pathways in vitro.
Cancer Research and Therapeutic Targeting: Elevated Galectin-3BP expression is correlated with tumor aggressiveness and poor prognosis in several cancers. It enhances galectin-3–mediated EGFR signaling, promoting epithelial-mesenchymal transition and metastasis. Recombinant Galectin-3BP is essential for studying these mechanisms and for screening inhibitors or antibodies that block its function.
Viral Infection Mechanisms: Galectin-3BP is upregulated in various viral infections (HIV-1, HCV, SARS-CoV-2) and can directly interact with viral components, limiting viral replication and influencing disease severity. Recombinant protein allows for controlled studies of these interactions and the development of prognostic assays.
Gene Therapy Vector Interactions: Galectin-3BP can aggregate and precipitate recombinant adeno-associated virus (rAAV) vectors, affecting their distribution and transduction efficiency in vivo. Using recombinant protein helps elucidate these interactions, optimizing gene delivery strategies.
Fibrosis and Inflammatory Disease Models: Its role in inducing pro-inflammatory cytokines and fibrosis, especially in severe COVID-19, makes recombinant Galectin-3BP useful for modeling fibrotic and inflammatory disease processes and testing anti-fibrotic interventions.
Diagnostic and Imaging Applications: Galectin-3BP is a promising biomarker for disease severity and progression, and its expression in tumors enables development of diagnostic imaging agents such as immunoPET tracers.
Standardization and Reproducibility: Recombinant protein ensures batch-to-batch consistency, defined purity, and absence of confounding serum factors, which are critical for reproducible experimental results.
Typical applications include:
Cell-based assays to study signaling, adhesion, and cytokine induction.
ELISA and immunoassays for biomarker validation.
Preclinical models for cancer metastasis and fibrosis.
Screening of inhibitors or antibodies targeting Galectin-3BP.
Mechanistic studies of viral infection and gene therapy vector biology.
In summary, recombinant Human Galectin-3BP is a versatile tool for investigating fundamental biological processes, disease mechanisms, and therapeutic strategies in immunology, oncology, virology, and regenerative medicine.
Recombinant Human Galectin-3BP can be used as a standard for quantification or calibration in ELISA assays, provided it is well-characterized, pure, and compatible with your assay system. This is a common practice in both commercial ELISA kits and custom assay development.
Supporting details:
Commercial ELISA kits for Galectin-3BP routinely use recombinant human Galectin-3BP as the standard for generating calibration curves. For example, the Quantikine ELISA kit is calibrated against a highly purified NS0-expressed recombinant human Galectin-3BP, and the assay recognizes both natural and recombinant forms, with parallel standard curves observed for both. This demonstrates that recombinant protein is suitable for quantification in these assays.
Standard preparation and use: The recombinant standard is typically reconstituted and serially diluted to generate a standard curve covering the assay’s dynamic range (e.g., 0.391–25 ng/mL or 156–10,000 pg/mL, depending on the kit). The absorbance values from these standards are used to interpolate the concentrations of unknown samples.
Assay compatibility: It is essential that the recombinant Galectin-3BP you use is full-length or matches the epitope(s) recognized by the capture and detection antibodies in your ELISA. Most commercial kits specify that their antibodies detect both natural and recombinant forms, but if you are developing your own assay or using a different recombinant construct (e.g., a fragment or tagged version), confirm that it is recognized equivalently.
Purity and activity: The recombinant standard should be of high purity (typically >80% by SDS-PAGE) and free from contaminants that could interfere with antibody binding or signal generation. Some recombinant proteins are not biologically active but are still suitable as quantitative standards if their epitopes are intact.
Validation: If you are substituting a recombinant standard for a native protein standard, it is good practice to validate that the standard curve generated is parallel to that obtained with native protein, ensuring accurate quantification.
Limitations and best practices:
Use the same buffer and diluent as recommended for your ELISA to avoid matrix effects.
Prepare fresh standards for each assay, as recombinant proteins can degrade or adsorb to plastic over time.
If your recombinant protein differs in glycosylation or structure from the native form, verify that this does not affect antibody recognition or quantification accuracy.
Summary: You can use recombinant human Galectin-3BP as a standard for ELISA quantification, as long as it is well-characterized, pure, and validated for your specific assay system. This approach is standard in both commercial and custom ELISA protocols.
Research Applications and Validation
Recombinant human Galectin-3BP (LGALS3BP) has been validated across multiple research applications in published studies:
Immunological Studies
In vitro research has demonstrated that Galectin-3BP stimulates natural killer cells and lymphokine-activated killer cell activity. The protein has been used to investigate immune modulation pathways, particularly in understanding how tumor-secreted factors influence immune cell function.
Cancer Biology Research
Galectin-3BP has been extensively studied in cancer research contexts. Knockdown and exogenous treatment studies have shown that Galectin-3BP is required for pancreatic ductal adenocarcinoma (PDAC) cell proliferation, migration, and invasion. Mechanistically, research has revealed that Galectin-3BP enhances galectin-3–mediated epidermal growth factor receptor signaling, leading to increased cMyc expression and epithelial-mesenchymal transition.
Tumor Microenvironment Studies
The protein has been validated for investigating tumor-stromal interactions. Published research has identified Galectin-3BP as a factor secreted by tumor cells that stimulates interleukin-6 expression in the bone marrow stroma and in the microenvironment of human neuroblastoma. Additionally, studies have demonstrated that Galectin-3BP secreted by breast cancer cells inhibits monocyte-derived fibrocyte differentiation.
Diagnostic and Therapeutic Development
Recombinant Galectin-3BP has been used as a standard in sandwich ELISA development kits for measuring natural and recombinant human Galectin-3BP in various biological samples. The protein has also served as a target antigen for developing antibody-based therapeutics, including antibody-drug conjugates for cancer treatment.
Bioassay Applications
The recombinant protein is validated for use in whole cell bioassays to assess functional interactions with binding partners including Galectin-3, Galectins-1 and -7, collagen types, fibronectin, beta-1 integrins, and nidogen.
To reconstitute and prepare Recombinant Human Galectin-3BP protein for cell culture experiments, follow these general best practices based on manufacturer recommendations and scientific protocols:
1. Reconstitution
Centrifuge the vial briefly before opening to ensure all lyophilized protein is at the bottom.
Reconstitute the protein in sterile, endotoxin-free water or sterile PBS (phosphate-buffered saline), depending on the manufacturer’s instructions. Typical reconstitution concentrations range from 0.1 to 1.0 mg/mL.
For example:
Some suppliers recommend 0.1–0.5 mg/mL in sterile water.
Others suggest 100 μg/mL in sterile PBS.
Gently mix the solution by pipetting up and down or swirling the vial. Avoid vigorous shaking or vortexing to prevent foaming and protein denaturation.
2. Sterilization
Filter sterilize the reconstituted protein solution using a 0.22 μm syringe filter in a clean bench or biosafety cabinet to ensure sterility for cell culture use.
3. Aliquoting
Aliquot the protein into small, single-use volumes to minimize repeated freeze-thaw cycles, which can degrade protein activity.
Store aliquots at –80°C for long-term storage. Avoid storing at –20°C if possible, as repeated freeze-thaw cycles can reduce activity.
4. Buffer Exchange (if needed)
If the protein is reconstituted in a buffer not compatible with your cell culture system (e.g., contains trehalose or mannitol), use an ultrafiltration device to exchange the buffer to your desired cell culture medium or PBS.
5. Endotoxin Removal (for sensitive cell types)
For experiments with primary cells or sensitive cell lines, consider removing endotoxins using a Detoxi-Gel column or similar method, especially if the protein will be used at high concentrations.
6. Quality Control
Check protein concentration using a suitable method (e.g., Bradford assay).
Verify endotoxin levels if required for your experiment (e.g., using a Limulus amebocyte lysate (LAL) assay).
7. Application in Cell Culture
Dilute the reconstituted protein to the desired working concentration in your cell culture medium immediately before use.
Avoid foaming during pipetting or mixing to maintain protein integrity.
Example Protocol:
Centrifuge the lyophilized protein vial briefly.
Reconstitute with sterile water or PBS to 0.1–1.0 mg/mL.
Gently mix and filter sterilize.
Aliquot and store at –80°C.
Thaw an aliquot, dilute to working concentration in cell culture medium, and add to cells.
Always refer to the specific product datasheet for any unique requirements or recommendations for your recombinant protein.
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
