Anti-Mouse CD8b.2 [Clone 53-5.8] — Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD8b.2 [Clone 53-5.8] — Purified in vivo PLATINUM™ Functional Grade

Product No.: C2836

[product_table name="All Top" skus="C2836"]

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Clone
53-5.8
Target
CD8b.2
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
Lyt-3.2, Ly-3.2
Isotype
Rat IgG1
Applications
Depletion
,
FA
,
FC
,
ICC
,
in vivo
,
WB

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
Mouse thymus or spleen
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% 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.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM™ antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
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
WB
FC The suggested concentration for this 53-5.8 antibody for staining cells in flow cytometry is ≤ 0.25 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
Additional Applications Reported In Literature ?
ICC
Depletion
FA
Additional Reported Applications For Relevant Conjugates ?
IF staining
IHC (Frozen)
IP


For specific conjugates of this clone, review literature for suggested application details.

Each investigator should determine their own optimal working dilution for specific applications.
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 53-5.8 recognizes an epitope on mouse CD8b.2.
Background
CD8 is made up of disulfide-linked α and β chains that form the α(CD8a)/β(CD8b) heterodimer and α/α homodimer. CD8 is part of the Ig superfamily that expresses primarily as CD8a homodimers. CD8a is a 32-34 kD type I glycoprotein that can also form heterodimers with CD8b. CD8 is an antigen co-receptor on T cells that mediates efficient cell to cell interactions within the immune system. CD8 coupled with the T cell receptor on the T lymphocyte recognizes an antigen displayed by an antigen presenting cell (APC) in the context of class I MHC molecules. The CD8 co-receptor also plays a role in T cell signaling by interacting with Lck (lymphocyte-specific protein tyrosine kinase) which leads to the activation of transcription factors that affect the expression of certain genes.
Antigen Distribution
CD8b.2 is expressed on the majority of thymocytes and cytotoxic T cell subsets.
Ligand/Receptor
MHC class I
NCBI Gene Bank ID
Research Area
Immunology

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 53-5.8 is most commonly used in vivo in mice to achieve selective depletion of CD8⁺ T cells by targeting the CD8β chain, which is a component of the CD8 coreceptor primarily expressed on cytotoxic T lymphocytes. This function enables researchers to study the roles of CD8⁺ T cells in immunity, infection, tumor surveillance, transplantation, and autoimmune disease models.

Key in vivo applications include:

  • CD8⁺ T cell depletion: Administration of clone 53-5.8 in mice results in robust elimination of peripheral CD8⁺ T cells, with high specificity for the CD8β chain, thereby sparing CD8⁺, CD11c⁺ dendritic cells.
  • Functional immune studies: By depleting CD8⁺ T cells, researchers can dissect the specific contribution of these cells to various immune responses, including responses to infection, vaccination, tumor challenge, and transplant acceptance or rejection.
  • Immunotherapy and memory cell research: Depletion with clone 53-5.8 allows comparison of the roles and functions of CD8⁺ T cells in primary or recall responses and is sometimes used to compare with CD8α depletion (which can have different immunologic effects).

Additional context:

  • Superiority for depletion: Evidence shows that clone 53-5.8 (anti-CD8β) is more efficient at depleting CD8⁺ T cells in vivo than anti-CD8α antibody clones.
  • Caveats: Residual CD8⁺ T cells that survive depletion can exhibit altered phenotypes and functions, possibly with enhanced cytotoxic capacity.
  • Alternative uses: While its main application is depletion in vivo, 53-5.8 is also used for in vitro blocking and immunofluorescence studies, though these are not in vivo contexts.

In summary, the principal in vivo use of clone 53-5.8 in mice is for selective CD8⁺ T cell depletion to enable precise functional studies of cell-mediated immunity.

Commonly used antibodies or proteins with 53-5.8 (anti-mouse CD8b.2) in the literature include key markers for defining T cell subsets and broader immune cell characterization. The most frequent companions are:

  • Anti-CD8α (commonly clone 53-6.7 or 53-6.72): Used to distinguish CD8a/b heterodimers from CD8a homodimers and for comprehensive CD8+ T cell analysis.
  • Anti-CD3: Identifies all T cells by targeting the T-cell receptor complex.
  • Anti-CD4: Labels helper T cell subsets, often to separate from cytotoxic CD8+ populations.
  • Anti-CD45: A pan-leukocyte marker, useful for general immune profiling or gating in flow cytometry.
  • Anti-CD11c: Sometimes used when characterizing dendritic cell populations, as some dendritic cells are CD8+.

These antibodies help distinguish CD8+ T cell subsets, segment T cell populations in flow cytometry, and support studies of cytotoxic cell functions. They are often combined in multicolor panels for murine immunophenotyping or T cell depletion experiments.

Clone 53-5.8 is a monoclonal antibody targeting mouse CD8β used extensively in immunology research for selective depletion of CD8+ T cells in vivo, with multiple studies highlighting its superior efficacy and diverse experimental outcomes.

Key findings from scientific literature citing clone 53-5.8 include:

  • High Depletion Efficiency: Clone 53-5.8 exhibits a high efficiency in depleting CD8+ T cells, outperforming other anti-CD8 monoclonal antibodies such as clone 53-6.72, making it a gold standard for mouse CD8+ T cell depletion.

  • Effects on Pulmonary Injury and Disease Models: In malaria models, administration of clone 53-5.8 reduced pulmonary vascular leakage and protected mice from experimental cerebral malaria (ECM), without affecting parasitemia levels. This demonstrates that the antibody can selectively mitigate CD8+ T cell-mediated tissue injury and improve survival during infection.

  • CD8+ T Cell Function Post-depletion: Residual CD8+ T cells, when present after clone 53-5.8-mediated depletion, may possess enhanced cytotoxic capability and altered surface marker expression, indicating effects on T cell function beyond mere depletion.

  • Impact on Differentiation and Phenotype: The use of clone 53-5.8 has facilitated functional studies revealing intrinsic requirements for CD8+ T cell differentiation, such as CD103-dependent pathways, and enabled precise phenotyping in competitive and adoptive transfer experiments.

  • Recommended Usage: Scientific and commercial sources recommend careful interpretation of experimental results given the antibody’s potency and specificity for functional CD8+ T cell removal.

  • Comparison with Other Clones: Peer-reviewed literature underscores that clone 53-5.8 is more selective and effective for targeting the CD8β subunit compared to some other clones against CD8α, which can spare non-cytotoxic CD8+ subsets.

In summary, citations of clone 53-5.8 focus on its high efficiency and specificity for depleting CD8+ T cells, its protective role in immune-mediated tissue injury in infectious disease models, and its utility in mechanistic studies of CD8+ T cell biology and differentiation.

Dosing regimens for the anti-mouse CD8β antibody clone 53-5.8 vary considerably across different experimental models, with variations in dose amounts, timing, and frequency depending on the specific research application and tumor model being studied.

Dose Amounts

The most commonly used dose is 5 mg/kg, which appears across multiple tumor models and experimental contexts. However, lower doses have also been employed successfully. In cancer models studying anthracycline resistance, researchers used 20 μg per mouse intraperitoneally, administered weekly starting from day 0. This represents a substantially lower dose compared to the standard 5 mg/kg regimen, yet still achieved effective CD8+ T cell depletion.

Timing and Frequency

The timing of antibody administration relative to tumor implantation shows significant variation. In some protocols, treatment begins the day before tumor implantation, continues on the day of implantation, and then weekly thereafter. This pre-emptive approach ensures CD8+ T cell depletion is established before tumor cells are introduced.

Other studies initiate treatment at different timepoints post-implantation. The specific timing depends on when investigators want to assess CD8+ T cell involvement in tumor control. The 5 mg/kg dose is typically administered on the day before, the day of implantation, and weekly thereafter in CD8 depletion studies.

For maintaining depletion throughout experiments, twice weekly dosing for 2-3 weeks is common in tumor models, while some protocols use weekly injections for long-term studies.

Model-Specific Considerations

Different tumor models employ distinct protocols. In CT26 and MC38 colon carcinoma models, as well as B16-F10 melanoma models, the standard 5 mg/kg dose is used with intraperitoneal administration. The duration of treatment varies from 2 weeks in MC38 models to 3 weeks in CT26 models.

Clone 53-5.8 is particularly valued because it completely depletes CD8+ T cells without depleting CD8+ CD11c+ dendritic cells, making it more selective than some alternative anti-CD8α antibodies. This selectivity allows for more precise interpretation of experimental results when distinguishing T cell effects from dendritic cell contributions.

References & Citations

Depletion
FA
Flow Cytometry
ICC
in vivo Protocol
General Western Blot Protocol

Certificate of Analysis

Formats Available

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