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

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

Product No.: C2843

[product_table name="All Top" skus="C2843"]

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Clone
I3/2.3
Target
CD45
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
T200, Ly-5, LCA
Isotype
Rat IgG2b κ
Applications
FC
,
IHC FF
,
IHC FFPE
,
in vivo
,
WB

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Data

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
Mouse lymphoma cell line
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
< 1.0 EU/mg as determined by the LAL method
Purity
≥95% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for this I3/2.3 antibody for staining cells in flow cytometry is ≤ 0.125 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
WB The suggested concentration for this I3/2.3 antibody for use in western blotting is 1-10 μg/ml.
Additional Applications Reported In Literature ?
IHC (Frozen)
IHC (Paraffin)
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Clone I3/2.3 recognizes an epitope on mouse CD45.
Background
CD45 is a 180-240kD glycoprotein member of the protein tyrosine phosphatase (PTP) family known for its involvement in regulating a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45 and its isoforms are vital regulators of T- and B-cell antigen receptor signaling. CD45 functions through its extracellular domain or through its cytoplasmic domain, and serves as a negative regulator of cytokine receptor signaling via JAK kinase supression. The large extracellular domain is highly glycosylated, and its multiple isoforms allow extensive variation in the structure of its side chains. CD45 isoforms show cell-type and differentiation-stage specific expression that can be used as markers that identify and distinguish between different types of immune cells. CD45R is an isoform of CD45 with a molecular weight of 220 kD. CD45R contains all three possible exons (A, B, and C); making it the longest protein generated from alternative splicing with a migration at 200 kD when isolated from T cells. Furthermore, B cells express CD45R with heavier glycosylation, bringing the molecular weight to 220 kD, hence the name B220. Notably, B220 expression is not only restricted to B cells and may also be expressed on activated T cells, on a subset of dendritic cells, and on other antigen-presenting cells. Additionally, activated and memory T lymphocytes express CD45RO which facilitates T cell activation. CD45RO lacks all three possible exons (A, B, and C), making it the shortest CD45 isoform.
Antigen Distribution
CD45 is expressed on all hematopoietic cells other than mature erythrocytes and platelets.
Ligand/Receptor
Galectin-1, CD2, CD3, CD4, TCR, CD22, Thy-1
Function
Phosphatase, T and B cell activation
NCBI Gene Bank ID
Research Area
Cell Biology
.
Immunology
.
Inhibitory Molecules
.
Neuroscience
.
Neuroscience Cell Markers
.
Stem Cell

Leinco Antibody 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.

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.

References & Citations

Flow Cytometry
IHC FF
IHC FFPE
in vivo Protocol
General Western Blot Protocol

Certificate of Analysis

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.