Recombinant Human ICAM-2 Fc Chimera

Recombinant Human ICAM-2 is a valuable tool for a wide range of research applications due to its well-characterized biological functions and versatility in experimental settings. Here are several key reasons why you should consider using Recombinant Human ICAM-2 in your research:

1. Study of Immune Cell Interactions

ICAM-2 is a critical adhesion molecule involved in immune responses. It mediates interactions between immune cells (such as lymphocytes, neutrophils, and NK cells) and endothelial cells, facilitating processes like:

• Lymphocyte recirculation
• Leukocyte rolling and firm adhesion
• Transendothelial migration (extravasation)
• NK-cell mediated clearance

Using recombinant ICAM-2 allows you to investigate these interactions in vitro, such as in cell adhesion assays, migration studies, or co-culture experiments.

2. Investigation of Cancer Cell Behavior

ICAM-2 has been shown to suppress cancer cell migration, invasion, and metastasis in various tumor types, including neuroblastoma and gastric cancer. Recombinant ICAM-2 can be used to:

• Study the mechanisms by which ICAM-2 inhibits tumor cell motility and invasion
• Evaluate its potential as a therapeutic target in cancer
• Perform functional assays (e.g., wound healing, Matrigel invasion) to assess the impact of ICAM-2 on cancer cell phenotype

3. Immuno-oncology and Tumor Microenvironment Studies

ICAM-2 plays a role in the tumor microenvironment by influencing immune cell recruitment and activity. Recombinant ICAM-2 can be used to:

• Investigate the effects of ICAM-2 on immune cell infiltration and anti-tumor immune responses
• Study the interaction between tumor cells and immune cells in co-culture systems
• Develop and test immunotherapeutic strategies targeting ICAM-2

4. Signal Transduction and Pathway Analysis

ICAM-2 is involved in signaling pathways that regulate immune cell activation and function. Recombinant ICAM-2 can be used to:

• Study the downstream signaling events triggered by ICAM-2 engagement
• Investigate the role of ICAM-2 in immune cell activation and cytokine production
• Perform biochemical assays to identify interacting partners and signaling molecules

5. High Purity and Consistency

Recombinant Human ICAM-2 is typically produced with high purity (>95% by SDS-PAGE) and is suitable for a variety of applications, including:

• Coating surfaces for cell adhesion and migration assays
• Use as a soluble protein in functional assays
• Immunoprecipitation and pull-down experiments

6. Versatile Applications

Recombinant ICAM-2 can be used in a variety of experimental formats, such as:

• Cell-based assays (adhesion, migration, invasion)
• ELISA and other immunoassays
• Protein-protein interaction studies
• In vitro and in vivo models of immune and cancer biology

7. Standardization and Reproducibility

Using recombinant ICAM-2 ensures consistency across experiments, reducing variability and improving the reproducibility of your results. This is particularly important for preclinical studies and therapeutic development.

Summary

Recombinant Human ICAM-2 is a powerful tool for studying immune cell interactions, cancer biology, and signal transduction. Its high purity, consistency, and versatility make it an excellent choice for a wide range of research applications, from basic science to translational and therapeutic studies.

Yes, recombinant human ICAM-2 can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated and matched to your assay system. Many commercial ELISA kits for human ICAM-2 use recombinant ICAM-2 as the standard, and these kits are designed to quantify both endogenous (natural) and recombinant forms of the protein.

Key considerations:

• Recognition by Antibodies: Most sandwich ELISA kits for ICAM-2 are validated to detect both endogenous and recombinant human ICAM-2, as the antibodies are raised against epitopes present on both forms. Always confirm that your assay’s capture and detection antibodies recognize the recombinant form you intend to use.
• Standard Curve Preparation: Prepare a standard curve with serial dilutions of the recombinant ICAM-2 in the same buffer or matrix as your samples to ensure accurate quantification. This is essential for reliable calibration.
• Protein Form and Purity: The recombinant ICAM-2 should be of high purity and, ideally, produced in a system that yields a protein with post-translational modifications similar to the native protein (e.g., glycosylation), as this can affect antibody recognition and assay performance.
• Unit Consistency: Ensure the concentration units of your recombinant standard (e.g., ng/mL, U/mL) match those used in your assay protocol and reporting.
• Validation: If you are substituting the standard provided in a commercial kit with your own recombinant protein, validate the standard curve for linearity, sensitivity, and parallelism with the kit’s original standard to ensure accurate quantification.

Summary Table: Use of Recombinant Human ICAM-2 as ELISA Standard

In summary: Recombinant human ICAM-2 is widely used as a standard in ELISA assays for quantification and calibration, but proper validation and matching to your assay system are essential for accurate results.

Recombinant Human ICAM-2 has been validated for several key applications in published research, primarily in studies of immune cell interactions, migration, and anti-tumor responses.

Validated Applications:

• Cell Adhesion and Migration Assays:
  Used to study lymphocyte and neutrophil interactions with endothelial cells, including crawling, polarization, and transendothelial migration (TEM) in vitro and in vivo. ICAM-2 supports neutrophil extravasation and monocyte locomotion, often via interaction with integrins such as LFA-1 and MAC-1.

• Immune Response and Surveillance Studies:
  Serves as a matrix protein to investigate antigen-specific immune responses, NK-cell mediated clearance, and lymphocyte recirculation. These assays often involve coating culture plates with recombinant ICAM-2 to facilitate cell adhesion and signaling.

• Anti-Tumor Immuno-Response Assays:
  Used as a culture matrix protein in vitro to study anti-tumor immune responses, including T cell and NK cell activation and cytotoxicity.

• Angiogenesis and Cellular Radioresistance:
  Expression and functional studies have linked ICAM-2 to angiogenesis and cellular radioresistance, often through its role in immune cell-endothelial interactions.

• Bioassays and ELISA Standards:
  Recombinant ICAM-2 is used as a bioactive ligand in cell-based bioassays and as a standard in ELISA to quantify ICAM-2 or study its binding properties.

Experimental Protocols:

• Plate Coating:
  Recombinant ICAM-2 is commonly coated onto culture plates (1–10 μg/well for 6-well plates) to facilitate cell adhesion and migration studies.

• Functional Blockade Studies:
  Used in experiments involving genetic knockout or antibody blockade to dissect the specific roles of ICAM-2 in leukocyte migration and immune cell dynamics.

Summary of Key Research Uses:

• In vitro: Cell adhesion, migration, and immune activation assays using coated recombinant ICAM-2.
• In vivo: Functional studies of neutrophil and lymphocyte extravasation, often in animal models with ICAM-2 blockade or deficiency.
• Immunological assays: ELISA, flow cytometry, and bioassays for quantifying ICAM-2 or its effects on immune cells.

These applications are supported by multiple published studies and product datasheets, confirming the utility of recombinant human ICAM-2 in immunology, cell biology, and cancer research.

Reconstitution Procedure

Recombinant Human ICAM-2 proteins are typically supplied in lyophilized form and require proper reconstitution before use. Begin by allowing the vial to reach room temperature, then reconstitute at a concentration of 0.4 mg/mL in sterile PBS (phosphate-buffered saline). Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation. Avoid vigorous shaking, as this can cause foaming and protein denaturation. Do not mix by vortexing or pipetting; instead, use gentle swirling motions to dissolve the lyophilized material.

Buffer Composition and Formulation

The reconstituted protein solution typically contains specific buffer components depending on the formulation. Standard formulations include 20 mM Tris-HCl buffer (pH 8.0), 0.15 M NaCl, 10% glycerol, and 1 mM DTT. Some formulations are supplied in PBS containing calcium and magnesium with sorbitol as a cryoprotectant. Always consult the product-specific datasheet for exact buffer composition, as formulations may vary between preparations.

Storage and Stability

After reconstitution, store the protein solution at 2-8°C for up to one week, or at -20°C to -70°C for longer-term storage (up to 3 months). Use a manual defrost freezer and minimize repeated freeze-thaw cycles, as these can compromise protein integrity and functionality. For extended storage beyond three months, maintain the lyophilized form desiccated at -20°C to -70°C, where it remains stable for six to twelve months.

Preparation for Cell Culture Applications

For cell culture experiments, dilute the reconstituted protein to your desired working concentration using serum-free medium or PBS. The final solution should be sufficiently dilute to ensure even coverage of the culture surface. For plate coating applications, use 1-10 µg of protein per well in a 6-well plate format. Incubate coated plates at 2-10°C overnight, then aspirate remaining material and allow plates to air dry or store at 2-8°C while maintaining sterility.

Important Considerations

Carrier-free versus carrier protein formulations: If your application requires carrier-free protein (without bovine serum albumin), use this formulation when BSA presence could interfere with your assay. For general cell culture applications, formulations with carrier protein may provide enhanced stability.

Optimal dilutions: Each laboratory should determine optimal dilutions for specific applications, as requirements vary based on cell type, culture conditions, and experimental design.

Centrifugation: Always centrifuge tubes before opening to ensure all lyophilized material is collected at the bottom of the vial.

Endotoxin testing: If using the protein in sensitive cell culture applications, verify endotoxin levels, as some formulations may require testing prior to use.