Anti-Mouse CD45 (Clone I3/2.3) – Purified in vivo GOLD™ Functional Grade

Clone I3/2.3 is primarily used in mice for immunological research applications centered around the identification, depletion, and tracking of leukocytes based on CD45 expression. CD45, also known as leukocyte common antigen (LCA), is a 180-240 kD glycoprotein present on all cells of hematopoietic origin except erythrocytes, making this antibody an essential tool for studying immune cell populations.

Primary In Vivo Applications

Leukocyte Depletion Studies represent a major application where the antibody is injected into mice to selectively deplete CD45-expressing immune cells. This allows researchers to study immune system function by removing specific cell populations and observing the resulting effects on disease models or immune responses.

Cell Trafficking and Migration Studies utilize clone I3/2.3 to track immune cell movement throughout the body. Since CD45 is expressed on nearly all immune cells, this antibody serves as a pan-leukocyte marker for monitoring immune cell distribution and migration patterns in various tissue compartments.

Immunological Characterization forms another critical application area where the antibody helps identify and characterize different immune cell populations within mouse models. Given that CD45 plays a key role in T cell receptor (TCR) and B cell receptor (BCR) signal transduction, this clone proves valuable for studying immune cell activation and function.

Technical Specifications for In Vivo Use

The antibody is manufactured specifically for in vivo applications with stringent quality controls. It contains less than 1.0 EU/mg endotoxin as determined by the LAL method, which is crucial for minimizing inflammatory responses that could confound experimental results. The purity exceeds 95% by SDS-PAGE and analytical SEC, ensuring minimal interference from contaminants. The antibody is typically formulated in phosphate buffered saline without carrier proteins, potassium, or preservatives, making it suitable for direct injection into mice.

The I3/2.3 anti-mouse CD45 antibody is commonly paired with several other antibodies and proteins in the literature to enable comprehensive characterization of immune cell populations in mouse models. These paired reagents serve different purposes depending on the experimental design.

T Cell and B Cell Markers

CD3 is one of the most frequently used antibodies alongside I3/2.3, serving as a T cell marker to identify and distinguish T lymphocyte populations within the CD45-positive leukocyte compartment.

B220 (a CD45 isoform) and other B cell markers are commonly used to identify B lymphocyte subsets within the overall leukocyte population defined by pan-CD45 staining.

Panel Design for Population Characterization

I3/2.3 is utilized in multi-parameter flow cytometry panels that allow researchers to either exclude or include specific cell populations. These panels typically incorporate several categories of markers:

Exclusion markers include viability dyes (live/dead discrimination), Ter119 (for erythrocyte exclusion), and markers for non-leukocyte cell types. These help remove unwanted populations from the analysis.

Inclusion markers combine CD45-positive gating with cell type-specific markers to identify particular leukocyte subsets of interest. This approach enables comprehensive characterization of the immune landscape in various disease models and research applications.

The strategic pairing of I3/2.3 with these complementary antibodies has made it a cornerstone tool in mouse immunology research, particularly for studies involving neuroinflammation, multiple sclerosis models, and investigations of microglia and macrophage function.

Clone I3/2.3 is a monoclonal antibody that specifically recognizes an epitope on mouse CD45, a protein crucial for regulating T- and B-cell activation, growth, and differentiation. The key findings from scientific literature utilizing this clone include:

1. Immune Phenotyping: Clone I3/2.3 is widely used for immune phenotyping in mouse models, particularly for identifying and analyzing CD45-expressing leukocytes, which are essential for understanding immune responses and disease modeling.

2. Reliability and Specificity: Over 10 peer-reviewed publications highlight the reliability and specificity of clone I3/2.3 in various applications, including flow cytometry, western blotting, and immunohistochemistry. For instance, it is commonly used to exclude or include specific leukocyte populations in studies.

3. Applicability in Research: The antibody is frequently cited in research related to immune cell characterization, making it a cornerstone tool in mouse immunology. It has been used in studies involving dendritic cells, multiple sclerosis models, and diabetes research.

4. Staining and Labeling: Clone I3/2.3 is used for staining cells in flow cytometry, with a suggested concentration of ≤ 0.125 μg per (10^6) cells. It is also utilized in western blotting and immunohistochemistry, though optimal concentrations may vary depending on the application.

Overall, clone I3/2.3 is a valuable resource for researchers studying immune cell biology and disease models in mice.

Based on the available search results, there is limited specific information about how dosing regimens of clone I3/2.3 vary across different mouse models. The search results primarily provide general usage guidelines rather than model-specific dosing protocols.

Flow Cytometry Applications

For flow cytometry applications, clone I3/2.3 (anti-mouse CD45) has a suggested concentration of ≤0.125 μg per 10⁶ cells in a volume of 100 μl. However, the manufacturer recommends that titration of the reagent is necessary for optimal performance for each specific application.

In Vivo Applications

While the search results mention that clone I3/2.3 is available in purified in vivo functional grade format, they do not provide detailed dosing regimens for specific mouse models. One reference notes that "subsequent doses were given every other day following initial loading dose" when using CD45 clone I3/2.3 in mouse studies, but this lacks specifics about the actual dose amounts or the particular mouse model used.

Western Blotting

For western blotting applications, the suggested concentration is 1-10 μg/ml, though this is not specific to in vivo dosing.

General Guidance

The available information emphasizes that each investigator should determine their own optimal working dilution for specific applications, suggesting that dosing varies considerably based on the experimental context, mouse strain, and specific research objectives. The lack of standardized dosing protocols across different mouse models in the search results indicates this remains an area where researchers must empirically optimize conditions for their particular experimental system.