Recombinant Mouse ICAM-2

Recombinant mouse ICAM-2 offers several significant advantages for research applications, particularly in studies involving cell adhesion, immune cell interactions, and inflammatory responses.

Key Research Applications

Cell Adhesion and Leukocyte Interactions

Recombinant mouse ICAM-2 serves as a functional substrate for studying neutrophil and leukocyte adhesion dynamics. When immobilized, it supports the adhesion of activated immune cells in a dose-dependent manner, making it ideal for investigating how adhesion molecules regulate immune cell behavior. This is particularly valuable for examining neutrophil crawling dynamics and transendothelial migration, as ICAM-2 plays a critical role in facilitating luminal interactions between neutrophils and endothelial cells.

Mechanistic Studies of Immune Responses

The protein enables detailed investigation of integrin-ligand interactions. ICAM-2 serves as a ligand for leukocyte adhesion proteins such as MAC-1 and LFA-1, allowing researchers to dissect the molecular mechanisms underlying immune cell recruitment and extravasation. This is essential for understanding how adhesion molecules regulate the stop-start crawling patterns and migration efficiency of immune cells.

Advantages of the Recombinant Format

Quality and Consistency

Recombinant proteins offer superior lot-to-lot consistency compared to traditional approaches. The recombinant format enables sequence verification and optimization, ensuring reproducible results across experiments and reducing experimental variability.

Functional Validation

Recombinant mouse ICAM-2-Fc chimera proteins have been functionally validated to support immune cell adhesion assays, providing confidence in their biological activity for your experimental design.

Diagnostic and Prognostic Potential

Beyond basic research, ICAM-2 measurements may have clinical relevance. Serum ICAM-2 levels show potential utility in diagnosis and monitoring of certain conditions, and ICAM-2 expression has been associated with favorable outcomes in immunotherapy contexts.

Using recombinant mouse ICAM-2 provides a well-characterized, reproducible tool for investigating fundamental mechanisms of immune cell adhesion and migration in your research.

Recombinant Mouse ICAM-2 protein can be used as a standard for quantification or calibration in ELISA assays, provided it is properly validated for this purpose. The key requirements are that the recombinant protein is of high purity, accurately quantified, and matches the native protein's immunoreactivity in your assay system.

Essential context and best practices:

• Standard Curve Preparation: ELISA quantification relies on a standard curve generated using known concentrations of the target protein. Recombinant Mouse ICAM-2 is commonly used as a standard in commercial ELISA kits for mouse ICAM-2, where its concentration is serially diluted to create the curve.
• Validation: It is critical to ensure that the recombinant ICAM-2 is recognized equivalently by the capture and detection antibodies used in your ELISA. Most commercial kits use recombinant ICAM-2 as their standard, confirming its suitability for quantification.
• Purity and Quantification: The recombinant standard should be highly purified and its concentration accurately determined, typically by absorbance at 280 nm or amino acid analysis. Impurities or inaccurate quantification can lead to errors in calibration.
• Matrix Effects: If your samples are in complex matrices (e.g., serum, plasma), consider preparing standards in the same matrix or using matrix-matched diluents to account for potential interference or recovery differences.
• Documentation: Refer to the ELISA kit protocol or guidelines for recommended standard preparation and dilution procedures.

Additional relevant information:

• Reactivity: Recombinant Mouse ICAM-2 is used in ELISA kits for quantifying ICAM-2 in mouse plasma, serum, and cell culture supernatants.
• Assay Range: Typical detection ranges for mouse ICAM-2 ELISA kits are from low picogram to nanogram per milliliter concentrations, depending on kit sensitivity.
• Controls: Internal controls and validation samples should be included to ensure assay accuracy and reproducibility.

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

In conclusion: You can use recombinant Mouse ICAM-2 as a standard for ELISA quantification, provided it is validated for your specific assay and prepared according to best practices for standard curve generation.

Recombinant Mouse ICAM-2 has been validated for several key applications in published research, primarily in studies of immune cell adhesion, migration, and signaling.

Validated Applications:

• Cell Adhesion Assays: Recombinant Mouse ICAM-2 has been used to support the adhesion of leukocyte cell lines (e.g., HL-60, HSB2) in vitro, demonstrating its functional activity in mediating integrin-dependent cell binding.
• Flow Cytometry: Antibodies against Mouse ICAM-2 are validated for flow cytometric analysis of cell surface expression, particularly on endothelial and hematopoietic cells.
• Immunohistochemistry (IHC): Recombinant Mouse ICAM-2 antibodies have been validated for IHC, enabling detection of ICAM-2 in tissue sections.
• Western Blot (WB): Recombinant Mouse ICAM-2 antibodies are validated for Western blotting to detect ICAM-2 protein in cell and tissue lysates.
• ELISA: Recombinant Mouse ICAM-2 is used as a standard or capture antigen in ELISA kits for quantitative measurement of ICAM-2 in plasma, serum, and cell culture supernatants.
• Blocking/Functional Studies: Recombinant Mouse ICAM-2 and its antibodies have been used in functional blockade experiments to study the role of ICAM-2 in neutrophil crawling, transendothelial migration (TEM), and immune cell interactions in vivo and in vitro.

Research Contexts:

• Neutrophil Migration and Extravasation: ICAM-2 is critical for neutrophil crawling and transmigration across endothelial barriers, as shown by genetic knockout and antibody blockade studies.
• Leukocyte-Endothelial Interactions: ICAM-2 mediates adhesion and migration of various immune cells (T cells, NK cells, monocytes) via integrin interactions, especially with LFA-1 and MAC-1.
• Angiogenesis and Tumor Biology: ICAM-2 expression influences angiogenesis, cellular radioresistance, and anti-tumor immune responses.
• Blood-Brain Barrier and Bronchial Epithelium Migration: ICAM-2 is involved in T cell migration across specialized barriers.

Summary Table:

These applications are supported by published research and product validation data, confirming the utility of recombinant Mouse ICAM-2 in immunology, cell biology, and translational studies.

Reconstitution of Lyophilized Recombinant Mouse ICAM-2

Recombinant Mouse ICAM-2 is typically supplied in lyophilized form and requires proper reconstitution before use in cell culture experiments. The reconstitution process should be performed aseptically to maintain protein integrity and sterility.

Reconstitution Procedure

Begin by briefly centrifuging the vial prior to opening to bring the contents to the bottom. Reconstitute the lyophilized protein using sterile, distilled water or sterile PBS, depending on the specific product formulation. The recommended reconstitution concentration is approximately 0.4 mg/mL in sterile PBS. However, it is not recommended to reconstitute to a concentration less than 100 μg/mL, as this may compromise protein stability.

Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation. Avoid vigorous shaking or vortexing, as this can cause foaming and protein denaturation. Instead, gently mix the solution by careful pipetting or gentle swirling.

Storage and Stability Considerations

Short-term Storage

Reconstituted protein solution can be stored at 4-8°C for 2-7 days. For longer-term storage, prepare aliquots of the reconstituted protein and store them at temperatures below -20°C, where they remain stable for approximately 3 months.

Long-term Storage

The lyophilized protein itself is stable for six to twelve months when stored desiccated at -20°C to -70°C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles, as these can significantly reduce protein activity and stability.

Carrier-Free Versus Carrier-Containing Formulations

When selecting a recombinant Mouse ICAM-2 product, consider whether a carrier-free or carrier-containing formulation is appropriate for your application. Carrier-free versions do not contain bovine serum albumin (BSA), making them suitable for applications where BSA could interfere with experimental results. In contrast, carrier-containing formulations with BSA are recommended for cell or tissue culture applications and as ELISA standards, as the carrier protein enhances stability and extends shelf-life.

Application-Specific Considerations

For cell adhesion assays, immobilized recombinant Mouse ICAM-2 supports the adhesion of activated cells in a dose-dependent manner. When coating plates with the protein, typical concentrations range from 12.5 μg/mL with 100 μL per well, which supports >70% cell adhesion after 1 hour incubation at 37°C. Optimal dilutions should be determined by your laboratory for each specific application, as requirements may vary based on experimental design and cell type.