Galectin-8, also known Gal-8 and PCTA-1, is a member of the Galectin family which selectively binds ß-galactoside residues. The galectins have been implicated in many essential functions including development, differentiation, cell-cell adhesion, cell-matrix interaction, growth regulation, apoptosis and RNA splicing. Galectins lack a typical signal peptide and can be localized to the cytosolic compartments where they have intracellular functions, however, via unknown, non-classical pathways, they can be secreted extracellularly. Gal-8 is expressed in liver, kidney, cardiac muscle, lung and brain. It is also widely expressed in tumoral tissues, such as lung and prostate carcinomas (1). It binds to a subset of cell surface integrins to modulate ECM-integrin interactions (2). As a soluble ligand, Gal-8 can inhibit cell adhesion, however, immoblized Gal-8 has been shown to promote cell adhesion (3). Alternatively spliced transcript variants encoding different isoforms have been identified (4).
The predicted molecular weight of Recombinant Human Galectin-8 is Mr 36 kDa.
Predicted Molecular Mass
36
Formulation
This recombinant protein was 0.2 µm filtered and lyophilized from modified Dulbecco’s phosphate buffered saline (1X PBS)and Betamercaptoethanol pH 7.2 – 7.3 with no calcium, magnesium, or preservatives.
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.
Leinco Protein Advisor
Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.
Recombinant Human Galectin-8 (rhGal-8) is a valuable tool for a wide range of research applications due to its well-characterized biological functions and versatility. Here are several key reasons why you should consider using recombinant human Galectin-8 in your research:
1. Functional Studies in Cell Biology
Trophoblast Invasion and Placental Development: rhGal-8 has been shown to stimulate adhesion, migration, and invasion of extravillous trophoblast (EVT) cells, making it essential for studies on placental development and pregnancy-related disorders. It upregulates matrix metalloproteinases (MMP-2 and MMP-9), which are critical for tissue remodeling and cell invasion processes.
Cell Adhesion and Migration: Galectin-8 modulates integrin-mediated signaling and extracellular matrix interactions, enabling studies on cell motility, wound healing, and tissue regeneration.
2. Immunology and Inflammation Research
Immune Cell Activation: rhGal-8 is a potent inducer of functional disease markers such as IL1B, TNF, IL6, MMP1, MMP3, and MMP13 in immune and inflammatory cells. It can be used to study cytokine production, immune cell activation, and inflammatory responses.
Autoimmune and Chronic Disease Models: Galectin-8 is implicated in autoimmune diseases (e.g., rheumatoid arthritis) and chronic inflammation, making it relevant for modeling disease mechanisms and screening therapeutic interventions.
3. Cancer and Tumor Biology
Regulation of Tumor Growth and Metastasis: Galectin-8 is highly expressed in certain cancers (e.g., lung and prostate carcinomas) and plays a role in tumor progression, metastasis, and cell survival. rhGal-8 can be used to investigate tumor cell behavior, signaling pathways, and potential therapeutic targets.
Matrix Remodeling: By modulating MMP expression, Galectin-8 influences the tumor microenvironment and extracellular matrix degradation, which are critical for cancer cell invasion and metastasis.
4. Host-Pathogen Interactions and Innate Immunity
Antimicrobial Activity: Galectin-8 exhibits antimicrobial activity against specific pathogens, including Streptococcus pneumoniae, and plays a role in host defense against molecular mimicry. This makes it useful for studies on innate immunity and host-pathogen interactions.
5. Standardization and Reproducibility
Consistent Quality: Recombinant proteins provide a consistent, highly purified source of Galectin-8, ensuring reproducibility across experiments and reducing batch-to-batch variability.
Flexibility in Experimental Design: rhGal-8 is suitable for a variety of assays, including cell culture, ELISA, Western blot, functional assays, and high-throughput screening.
6. Mechanistic and Pathway Studies
Signaling Pathway Analysis: rhGal-8 can be used to dissect signaling pathways involving NF-κB, integrins, and MMPs, providing insights into cellular responses and regulatory mechanisms.
7. Therapeutic and Diagnostic Development
Drug Screening: Galectin-8 is a potential target for therapeutic intervention in cancer, autoimmune diseases, and inflammatory disorders. rhGal-8 can be used in drug screening and target validation studies.
Biomarker Research: Due to its involvement in disease progression, Galectin-8 is a candidate biomarker for various conditions, and recombinant protein can be used in assay development and validation.
In summary, recombinant human Galectin-8 is a versatile and well-characterized protein that enables robust and reproducible research in cell biology, immunology, cancer, and infectious disease. Its ability to modulate key cellular processes and signaling pathways makes it an essential reagent for advancing scientific understanding and developing new therapeutic strategies.
Recombinant Human Galectin-8 can be used as a standard for quantification or calibration in ELISA assays, provided it is compatible with your assay system and properly validated.
Key considerations and supporting details:
ELISA Standards: Many commercial ELISA kits for human Galectin-8 use recombinant Galectin-8 as their reference standard for generating calibration curves. This is a common practice, as recombinant proteins offer high purity and defined concentration, which are essential for accurate quantification.
Compatibility: It is crucial to confirm that your ELISA antibodies recognize both recombinant and native Galectin-8 with similar affinity. Most sandwich ELISA kits are designed to detect both forms, but some kits specify detection of only native or recombinant protein. Always check your kit’s documentation for compatibility.
Standard Preparation: Prepare the recombinant Galectin-8 standard according to your ELISA protocol, typically by serial dilution in the recommended diluent to cover the assay’s dynamic range. Standards should be freshly prepared for each assay to ensure accuracy.
Validation: Validate the use of recombinant Galectin-8 as a standard in your specific assay by confirming parallelism between the standard curve and sample dilution curves. This ensures that matrix effects or differences in glycosylation do not affect quantification.
Formulation: Recombinant Galectin-8 is often supplied with carrier proteins (e.g., BSA) to enhance stability and solubility, which is suitable for use as an ELISA standard. Ensure your standard matches the formulation recommended for your assay.
Documentation: Some protocols and product datasheets explicitly recommend recombinant Galectin-8 for use as an ELISA standard. For example, “In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard”.
Summary of best practices:
Use recombinant Galectin-8 of known concentration and purity.
Confirm antibody compatibility with recombinant protein.
Prepare standards according to your ELISA kit’s instructions.
Validate standard curve performance in your assay matrix.
If your ELISA kit or protocol specifies detection of only native Galectin-8, or if there are concerns about post-translational modifications affecting antibody recognition, additional validation may be required. Otherwise, recombinant Human Galectin-8 is widely accepted as a standard for ELISA quantification.
Recombinant Human Galectin-8 has been validated for a range of applications in published research, primarily in functional cell-based assays and molecular studies. Key validated applications include:
Cell adhesion, migration, and invasion assays: Recombinant human Galectin-8 has been used to stimulate and assess the adhesion, migration, and invasion of human trophoblast cell lines (HTR-8/SVneo) and primary cytotrophoblasts, demonstrating its role in modulating these cellular processes relevant to placental development and cancer metastasis.
Gene and protein expression analysis: Its effects on the expression of matrix metalloproteinases (MMP-2, MMP-9) and integrin subunits have been quantified using qPCR, cell-based ELISA, and gelatin zymography.
Functional disease marker induction: In osteoarthritis research, recombinant Galectin-8 has been used to treat human chondrocyte cultures, leading to the induction of inflammatory and matrix-degrading markers (e.g., IL1B, TNF, IL6, MMP1, MMP3, MMP13) and downregulation of cartilage matrix genes (COL2A1, ACAN), with effects measured at both mRNA and protein levels.
Cell adhesion modulation in cancer: It has been applied to study the adhesion of multiple myeloma cells to vascular endothelium, showing enhancement of cell adhesion under dynamic flow conditions.
Hemagglutination and cell adhesion inhibition: Recombinant Galectin-8 has been validated in hemagglutination assays (agglutination of human red blood cells) and in assays measuring inhibition of cell-substrate adhesion, particularly in hepatocarcinoma cell lines.
Immunoassays and ELISA: Recombinant Galectin-8 is commonly used as a standard or functional ligand in ELISA and other immunoassays to study protein-protein or protein-carbohydrate interactions.
Summary Table: Validated Applications of Recombinant Human Galectin-8
Application Type
Example/Details
Reference
Cell adhesion, migration, invasion
Trophoblast and cancer cell lines; invasion and migration assays
Gene/protein expression analysis
qPCR, ELISA, zymography for MMPs, integrins, cytokines
Disease marker induction
Osteoarthritis chondrocyte cultures; inflammatory and matrix markers
Hemagglutination
Agglutination of human RBCs; carbohydrate-binding specificity
Cell adhesion inhibition
Hepatocarcinoma cell lines; inhibition of cell-substrate adhesion
Immunoassays/ELISA
Used as standard or ligand in ELISA and related immunoassays
Additional Notes:
Recombinant human Galectin-8 is also widely used in studies of cancer biology, immune regulation, and inflammation, often as a tool to dissect its role in cell signaling, tumor progression, and immune cell function.
The protein is typically validated for use in in vitro cell culture systems, molecular assays, and sometimes as a reference standard in biochemical assays.
If you require details on a specific application or protocol, please specify the experimental context.
To reconstitute and prepare Recombinant Human Galectin-8 protein for cell culture experiments, dissolve the lyophilized protein in sterile distilled water or PBS to a concentration not less than 100 μg/mL. Incubate the solution at room temperature for at least 20 minutes to ensure complete dissolution, and do not vortex—gentle mixing is preferred to preserve biological activity.
Step-by-step protocol:
Briefly centrifuge the vial before opening to collect the powder at the bottom.
Add sterile distilled water or PBS to achieve a concentration ≥100 μg/mL.
Gently mix by pipetting or slow inversion; avoid vigorous shaking or vortexing.
Incubate at room temperature for 20 minutes to ensure full reconstitution.
For long-term stability or to prevent adsorption, add a carrier protein such as 0.1% BSA or 10% FBS if required for your application.
After reconstitution, aliquot the solution to avoid repeated freeze-thaw cycles and store at 2–8 °C for up to 1 week, or at −20 °C or below for longer-term storage.
For cell culture experiments, further dilute the stock solution in your desired culture medium or buffer, ensuring compatibility with your assay conditions.
Additional notes:
If endotoxin sensitivity is critical for your cell culture, confirm the endotoxin level is <1.0 EU/μg protein.
Always use sterile technique to prevent contamination.
Avoid concentrations below 100 μg/mL during initial reconstitution to maintain protein stability.
This protocol ensures optimal solubility and biological activity of recombinant Galectin-8 for cell-based assays.
References & Citations
1. Nagy, N. et al. (2002) Gut. 50:392
2. Cárcamo, C. et al. (2006) Exp. Cell Res. 312:374
3. Levy, Y. et al. (2001) J. Biol. Chem. 276:31285
4. Bidon, N. et al. (2001) Gene. 274:253
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
