Anti-Mouse MHC Class I (H-2Kk, H-2Dk) (Clone 15-3-1S) – Purified in vivo GOLD™ Functional Grade

The clone 15-3-1S is a monoclonal antibody that reacts with the mouse H-2Kk and H-2Dk MHC Class I alloantigens. This antibody is used in in vivo mouse studies primarily for its ability to target and interact with MHC Class I molecules, which are crucial for presenting antigens to CD8 T cells. MHC Class I molecules are expressed on the surface of nearly all nucleated cells and play a significant role in the immune system by displaying peptides from inside the cells to cytotoxic T cells.

In in vivo administration, the clone 15-3-1S can be used to study immune responses and modulate the function of MHC Class I molecules. For example, it might be employed to block or analyze the MHC Class I interaction with T cells, which is important in both normal immune function and in disease models, such as cancer or viral infections.

The 15-3-1S monoclonal antibody is applicable in various research settings, including:

• Flow cytometry: To analyze the expression of MHC Class I molecules on different cell types, helping in understanding immune cell interactions.
• Immunomodulation studies: It can be used to modulate immune responses by blocking or enhancing the interaction between MHC Class I and CD8 T cells.

Overall, the clone 15-3-1S is a valuable tool in research settings where understanding or manipulating MHC Class I molecules is needed.

Specifications and Usage:

• Isotype: Mouse IgG2a, ?
• Recommended Dilution Buffer: InVivoPure pH 7.0 Dilution Buffer
• Conjugation: Unconjugated but can be conjugated as needed
• Formulation: PBS, pH 7.0
• Purity: >95% as determined by SDS-PAGE.

The correct storage temperature for sterile packaged biological products, such as antibodies or clones, can vary depending on the specific requirements of the product. However, for many types of biological samples, including antibodies, short-term storage (up to 3 months) is typically recommended at 2-8°C in a refrigerator. For long-term storage, it is often advised to store the product at -20°C to prevent degradation and maintain stability, especially after opening or reconstitution.

Without specific instructions for the clone 15-3-1S, it would be prudent to follow general guidelines for similar products. Always check the product datasheet or manufacturer's instructions for specific storage recommendations.

The most commonly used antibodies or proteins alongside 15-3-1S (typically referring to antibodies targeting the CA 15-3 antigen/MUC1) in the literature include:

• Monoclonal antibodies such as mouse IgG clones (e.g., Clone A and B) specifically targeting CA 15-3.
• Polyclonal antibodies, including avian IgY antibodies generated against CA 15-3 epitopes.
• Human IgG antibodies recognizing peptide epitopes of the CA 15-3 antigen, often with specificity for varied posttranslational modifications.
• MUC1 proteins, frequently used as antigens; assays often utilize MUC1 derived from breast cancer cell lines or recombinant MUC1 constructs as capture/detection reagents.
• Control antibodies for experimental validation, such as isotype control antibodies (e.g., anti-tetanus toxoid, or non-specific IgG).
• Other monoclonal antibodies or fusion proteins conjugated to detection tags (for example, mouse monoclonal detection antibodies labeled with sulfo-tag for ECL assays).

Common combinations in diagnostic or research settings include:

• Anti-CA 15-3 monoclonal antibodies (mouse IgG, clones A and B) and anti-CA 15-3 IgY polyclonal antibodies: These are used for comparative sensitivity/reactivity in ELISA platforms.
• CA 15-3 antigen (MUC1) and detection anti-human IgG antibodies: Used for quantifying autoantibodies against CA 15-3 in human serum using immunoassays.
• Different capture and detection antibody pairs: Often combining monoclonal and polyclonal antibodies to increase assay coverage of native and modified MUC1 epitopes.

In summary, other widely used antibodies or proteins with 15-3-1S targets include mouse monoclonal IgG anti-CA 15-3 antibodies (multiple clones), avian IgY polyclonal anti-CA 15-3 antibodies, human IgG detection antibodies, and recombinant or cell line–derived MUC1/CA 15-3 proteins as assay reagents. Control antibodies and isotype controls are also regularly used in these studies for specificity and validation.

Key findings from scientific literature regarding HL-60 clone 15 (often termed HC15 or clone 15-3-1S) focus on its use as a model cell line for studying eosinophil biology and leukocyte migration, especially as an alternative to primary eosinophil isolation.

Main findings include:

• Differentiation Potential: HL-60 clone 15 can be differentiated into eosinophil-like cells using sodium butyrate, with or without interleukin-5 (IL-5), making it a versatile model for eosinophil research where primary cell availability is limited.
• Proteomic and Phenotypic Characterization: Proteomic analyses show differentiated clone 15 cells share considerable similarities with eosinophils (Eos), particularly in granule protein expression (e.g., lactotransferrin, eosinophil cationic protein). However, some aspects of their protein profile and function remain more similar to neutrophils (Neutros), depending on the differentiation state.
• Surface Marker Expression: Upon differentiation, the HL-60 clone 15 line upregulates critical eosinophil markers such as GATA-1, IL-5R, EMR1, and TREM-1.
• Secretory and Inflammatory Features: Differentiated HC15 cells secrete key inflammatory mediators, including CCL-5, EPX (eosinophil peroxidase), and IL-8. The presence of IL-5 in differentiation protocols enhances this secretory profile.
• Functional Comparisons: Although these cells display weaker activation responses than primary eosinophils, their migration and adherence patterns to endothelial cells are similar, supporting their utility for functional studies of leukocyte migration and inflammation.
• Usefulness in Eosinophil Research: The ease of culture and differentiation of HL-60 clone 15 (relative to isolating primary eosinophils) positions it as a practical surrogate for mechanistic, proteomic, and functional studies of eosinophil and innate immune cell biology.

These findings collectively underscore HL-60 clone 15's value as a standardized, accessible experimental system for dissecting eosinophil-mediated processes in immunity and inflammatory disease, while also highlighting some limitations relative to primary human cells.