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

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

Product No.: C2832

[product_table name="All Top" skus="C2832"]

- -
- -
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 κ
Applications
Depletion
,
FA
,
FC
,
ICC
,
in vivo
,
WB

- -
- -
Select Product Size
- -
- -

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
< 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
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.

The clone 53-5.8 is an anti-mouse CD8? (Lyt 3.2) monoclonal antibody that serves as a powerful tool for CD8+ T cell depletion in in vivo mouse studies. This rat IgG1 antibody specifically targets the ? chain of the CD8 differentiation antigen on mouse cells.

Primary Applications in In Vivo Studies

CD8+ T Cell DepletionThe 53-5.8 antibody is primarily used to deplete CD8+ T cells in living mice for experimental purposes. When administered in vivo, this antibody has been shown to completely deplete CD8+ T cells while notably preserving other cell populations. Importantly, it does not deplete CD8+ CD11c+ dendritic cells, making it a selective tool for targeting specific immune cell subsets.

Immunological ResearchResearchers utilize this antibody in studies investigating the role of conventional CD8??+ T cells in various disease models and immune responses. The depletion approach allows scientists to examine what happens when CD8+ T cells are removed from the immune system, helping to understand their contribution to immune responses, tumor immunity, and autoimmune conditions.

Mechanism and Specificity

The 53-5.8 clone recognizes an epitope on mouse CD8b.2 and is specific for Ly-3.2 phenotype, showing only low reactivity with Ly-3.1 strains such as AKR, C58, MRL, and PL mice. This specificity is important because CD8 can exist as either a heterodimer (CD8??) or homodimer (CD8??), and this antibody specifically targets the ? chain component.

Commercial Availability and Formats

The 53-5.8 clone is widely available from multiple suppliers in various formats optimized for in vivo use, including low endotoxin formulations specifically designed for animal studies. It's available in bulk quantities ranging from milligrams to multi-gram amounts, reflecting its common use in research. The antibody is also available in different conjugated forms for flow cytometry applications to monitor depletion efficiency.

This antibody has become a standard tool in immunological research, with over 100 product citations, demonstrating its widespread acceptance and reliability in the scientific community.

The recommended storage temperature for sterile packaged clone 53-5.8 (anti-mouse CD8b.2 antibody) is 2–8°C for short-term storage, typically up to one month. For longer-term storage, aliquoting and freezing at lower temperatures (such as -20°C or -80°C) may be advised, depending on specific product guidance.

Key details:

  • Short-term storage (up to 1 month): Store undiluted and sterile at 2–8°C.
  • Longer-term storage: To preserve activity for extended periods, aliquot under sterile conditions and freeze at -20°C or -80°C.
  • Do not freeze liquid antibody for routine storage at 2–8°C, as freeze-thaw cycles can reduce activity.
  • Protect from light if labeled with a fluorochrome such as PE or PerCP/Cyanine5.5.
  • The product is usually formulated in phosphate-buffered saline without carrier proteins or preservatives, so avoid repeated freeze-thaw cycles by aliquoting.

For maximum stability and shelf life, always follow manufacturer-specific recommendations on the datasheet supplied with the antibody batch.

Commonly Used Antibodies and Proteins with 53-5.8

The 53-5.8 monoclonal antibody specifically targets the mouse CD8? chain (also known as Ly-3.2). Researchers often combine 53-5.8 with other antibodies and proteins to study CD8+ T cell biology, immune depletion, and cellular interactions. Here are some of the most commonly used reagents co-employed with 53-5.8 in the literature:

Anti-CD8? Antibodies

  • 53-6.72: Frequently used alongside 53-5.8 to target the CD8? chain, enabling comprehensive coverage of the CD8 co-receptor (CD8?? heterodimer) on cytotoxic T cells and thymocytes.
  • Other CD8? clones: Depending on the experimental context, additional anti-CD8? clones may be used, but 53-6.72 is most commonly paired with 53-5.8 for depletion or surface staining.

Additional T Cell Markers

  • Anti-CD3: Used to pan-activate or identify T cells.
  • Anti-CD4: For identifying and comparing CD4+ helper T cell populations.
  • Anti-CD25: Used in functional studies, especially for regulatory T cell characterization.
  • Anti-CD44, Anti-CD62L: For identifying memory and naive T cell subsets.

Depletion and Functional Studies

  • Anti-CD11c: Sometimes used to distinguish CD8+ T cells from CD8+ CD11c+ dendritic cells, since 53-5.8 does not deplete dendritic cells.
  • Anti-thymocyte markers: For studying T cell development in the thymus, as both CD8? and ? chains are expressed early.

Functional Assays and Recombinant Proteins

  • MHC class I tetramers: To assess antigen-specific CD8+ T cell responses, often in combination with FACS staining using 53-5.8.
  • Anti-IFN?, Anti-TNF?: For cytokine profiling of CD8+ T cell effector function.
  • Anti-Granzyme B, Anti-Perforin: To evaluate cytotoxic activity.

Isotype Controls

  • Rat IgG1: Used as an isotype control for 53-5.8 (which is a rat IgG1 antibody).
  • Rat IgG2a: Isotype control for anti-CD8? (53-6.7, rat IgG2a).

Typical Experimental Combinations

A standard experimental setup might involve:

  • Surface staining: 53-5.8 (CD8?), 53-6.72 (CD8?), CD3, CD4, CD44, CD62L for basic T cell subset analysis.
  • Depletion experiments: 53-5.8 alone or with 53-6.72 for selective depletion of CD8+ T cells in vivo.
  • Intracellular cytokine staining: 53-5.8 for CD8?, coupled with antibodies against cytokines such as IFN?, TNF?, Granzyme B.

Summary Table

TargetCommon Antibody Clone(s)Purpose/Context
CD8? (Ly-3.2)53-5.8Core reagent, depletion, staining
CD8?53-6.72Co-staining/depletion with CD8?
MHC class IVarious tetramersAntigen-specific T cell analysis
CD3, CD4, CD25Various clonesT cell subset identification
CD44, CD62LVarious clonesMemory/naive T cell profiling
CD11cVarious clonesDendritic cell distinction
IFN?, TNF?, Granzyme BVarious clonesEffector function assessment
Rat IgG1, IgG2aN/AIsotype controls

Key Literature Context

  • 53-5.8 is most often paired with 53-6.72 for comprehensive CD8+ T cell studies in mice, especially in depletion, functional assays, and multiparameter flow cytometry.
  • When targeting CD8+ subsets, antibodies against activation markers (CD44, CD62L), cytokines (IFN?, TNF?), and cytotoxic molecules (Granzyme B) are frequently combined with 53-5.8 to profile T cell function.
  • Dendritic cell distinction is important, as 53-5.8 does not deplete CD8+ CD11c+ dendritic cells.
  • Isotype controls are essential, with rat IgG1 for 53-5.8 and rat IgG2a for 53-6.72.

These combinations reflect the standard toolkit for studying CD8+ T cell biology in the mouse system, leveraging 53-5.8 as a key reagent for identifying, depleting, or functionally analyzing CD8?-expressing cells.

Key findings from scientific literature citing clone 53-5.8, an anti-mouse CD8? monoclonal antibody, include its effectiveness for depleting CD8+ T cells in mice, the distinct effects it has on immune cell populations compared to anti-CD8? antibodies, and its utility in dissecting CD8+ T-cell–mediated immune functions and pathologies.

Essential findings and context from major citations:

  • Specificity and Potency: Clone 53-5.8 targets the ?-chain (CD8?) of the mouse CD8 antigen, which is expressed predominantly on CD8+ T cells. Compared to anti-CD8? clones (e.g., 53-6.7), clone 53-5.8 more efficiently depletes CD8+ T cells in vivo and is recommended when maximal depletion is needed.

  • Functional Implications of Depletion: Depletion using clone 53-5.8 leaves behind a population of CD8+ T cells with distinct functional and phenotypic properties. Surviving cells after anti-CD8? treatment may retain or even exhibit increased cytotoxic function, especially during primary immune responses. This has major implications for experiments analyzing the absence or function of CD8+ T cells as results can be confounded by the characteristics of the surviving population.

  • Interpretation Challenges: The 53-5.8 antibody induces substantial CD8 internalization, complicating detection by flow cytometry post-depletion (staining intensity drops), and can make proper identification of residual CD8+ T cells difficult without careful gating strategies.

  • Pathophysiological Applications: Clone 53-5.8 has been instrumental in demonstrating the pathogenic roles of CD8+ T cells in disease models. For instance, in a cerebral malaria mouse model, anti-CD8? (53-5.8) treatment depleted CD8+ T cells, reduced pulmonary vascular leakage, and improved survival, without affecting parasite burden—demonstrating the direct pathogenic role of CD8+ T cells in pulmonary complications.

  • Comparisons and Limitations: Combining clone 53-5.8 with anti-CD8? mAbs does not increase depletion efficiency beyond that achieved with clone 53-5.8 alone, and using less effective clones (like anti-CD8? 53-6.7) can result in survival of memory-type T cells, potentially skewing long-term immunological studies.

Summary Table: Clone 53-5.8 vs. Anti-CD8? (53-6.7)

FeatureClone 53-5.8 (Anti-CD8?)Clone 53-6.7 (Anti-CD8?)
Depletion efficacyHigherLower
Surviving cell stateRetain/increase cytotoxicityMore central memory-like
Flow cytometry detectionReduced due to CD8 internalizationLess reduction
Use in dual depletionNo increased efficacyN/A

Additional Notes:

  • Clone 53-5.8 is referenced as a gold standard tool for functional CD8+ T cell depletion in mouse immunology experiments.
  • Researchers are advised to interpret results cautiously, considering possible residual and altered CD8+ populations after depletion.

In summary, the scientific literature characterizes clone 53-5.8 as a potent, widely used tool for mouse CD8+ T-cell depletion that profoundly influences subsequent immune responses and experimental outcomes.

References & Citations

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

Certificate of Analysis

- -
- -

Formats Available

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