Anti-Mouse CD8a (Ly 2.2) [Clone 2.43] — Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD8a (Ly 2.2) [Clone 2.43] — Purified in vivo PLATINUM™ Functional Grade

Product No.: C2837

[product_table name="All Top" skus="C2837"]

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Clone
2.43
Target
CD8a
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
Ly-2, Ly-35, Ly-B, Lyt-2, Lyt2, Ly 2.2
Isotype
Rat IgG2b
Applications
Depletion
,
FA
,
FC
,
ICC
,
IF Staining
,
IHC FFPE
,
in vivo
,
IP

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
Mouse CTL clone L3
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 GOLD™ 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
FC The suggested concentration for this 2.43 antibody for staining cells in flow cytometry is ≤ 1.0 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 ?
Depletion
FA
Additional Reported Applications For Relevant Conjugates ?
ICC
IF Staining
IHC (Paraffin)
IP
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 2.43 recognizes an epitope on mouse CD8a.
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
CD8a is present on the surface of most thymocytes and a subpopulation of mature T-lymphocytes which include most T suppressor/cytotoxic-cells.
NCBI Gene Bank ID

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 2.43 is a rat anti-mouse CD8? monoclonal antibody used in in vivo mouse studies to selectively deplete CD8+ T cells. Researchers administer clone 2.43 to mice, most commonly via intraperitoneal or intravenous injection, at doses typically ranging from 100??g to 500??g per mouse, with 250??g per mouse frequently used as a standard for effective depletion.

Context and Key Details:

  • Target and Specificity: Clone 2.43 binds specifically to the CD8? chain of the mouse CD8 protein, a surface marker found on CD8+ T cells, which are crucial for cell-mediated immune responses.
  • Research Purpose: The primary use of clone 2.43 is to create a model in which CD8+ T cells are acutely or chronically ablated, facilitating the study of their role in immunological processes, disease models (such as tumor immunity or infection), and therapeutic interventions.
  • Administration: The antibody is supplied in low-endotoxin, in vivo-grade preparations to minimize adverse reactions. It is typically administered at 100–500??g per mouse, with routes and frequency tailored to study design. Single or repeated dosing can be used, but repeated administration may elicit immune responses against the rat antibody.
  • Controls: Experiments often include isotype controls or untreated groups to distinguish effects due to CD8+ T-cell depletion from other possible antibody-driven effects.

Advantages:

  • High specificity: Efficient and selective depletion of CD8+ T cells with minimal impact on other immune populations.
  • Well-established: Widely validated in diverse mouse models and research contexts.

Potential Limitations:

  • Off-target or immunomodulatory effects may occur, although these are generally limited due to high specificity.
  • Immunogenicity: The rat origin of the antibody can induce anti-rat immune responses on repeated dosing, influencing longer term studies.

Additional Applications:

  • Clone 2.43 has served as a source of sequence for engineered derivatives (e.g., minibodies) in imaging and mechanistic studies.

In summary, clone 2.43 is the gold standard antibody for in vivo depletion of CD8+ T cells in mouse models, crucial for dissecting CD8+ T cell contributions in immunological research.

The correct storage temperature for sterile packaged clone 2.43 (anti-mouse CD8? monoclonal antibody) depends on the intended storage duration:

  • Short-term (up to 1-2 weeks): Store at 2–8°C or 4°C as supplied.
  • Medium-term (up to 2 months): Store at 2–8°C, according to some suppliers.
  • Long-term (up to 12 months): Store at -20°C.
  • Very long-term (over 12 months): Store at -80°C.

For all durations:

  • Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • Do not dilute prior to aliquoting for long-term storage.
  • Store undiluted solution at 4°C in the dark for short term.

Summary Table: Storage Duration vs. Temperature

Storage DurationTemperature
Short term (1-2 weeks)4°C (2–8°C)
Medium term (up to 2 months)2–8°C
Long term (12 months)-20°C
Very long term (>12 months)-80°C

Always consult the product datasheet or supplier for specific recommendations about your exact formulation and packaging.

Commonly used antibodies or proteins with 2.43 (anti-mouse CD8?, clone 2.43) in the literature are typically selected to identify and characterize different leukocyte populations or functional states in mouse immunology experiments. The most relevant co-used antibodies/proteins include:

  • Anti-CD4 (e.g., clone GK1.5): For distinguishing and depleting CD4+ T cells versus CD8+ T cells in phenotyping or depletion studies.
  • Anti-CD3: A pan-T cell marker, commonly used alongside anti-CD8? to identify all T cells.
  • Anti-CD8? (e.g., clone 53-5.8 or 53-6.7): For more refined phenotyping of CD8+ T cells specifically expressing the ? chain.
  • Anti-B220/CD45R: Marks B cells and some subsets of T cells; often used to exclude non-T cells in flow cytometry.
  • Anti-NK1.1 or DX5: Used to distinguish NK cells from T cells, especially in studies of cytotoxic lymphocyte populations.
  • Anti-MHC class I and II (e.g., I-A/I-E, H-2K^b/D^b): For context about antigen presentation and to characterize APCs in conjunction with CD8+ T cell studies.

Other reagents frequently employed:

  • Isotype controls: Such as rat IgG2b, to control for non-specific binding when using clone 2.43 (which is a rat IgG2b).
  • Viability dyes: To gate out dead cells during flow cytometry, critical in multi-color panels with 2.43.

Common applications in which these markers are used together include:

  • Flow cytometry (FACS), including multicolor panels
  • Cell depletion in vivo or in vitro
  • Immunoprecipitation and immunofluorescence

Panel design and antibody choice depend on experimental goals, such as distinguishing T cell subsets, characterizing effector/memory populations, or confirming depletion efficacy.

Summary table: Key antibodies and their use with 2.43

Marker/AntibodyTypical Purpose
CD4 (GK1.5)Identify, quantify, or deplete CD4+ T cells
CD3Identify all T cells
CD8? (e.g., 53-6.7)Quantify CD8+ T cell subsets, distinguish from other CD8+ cells
B220/CD45RExclude B cells and non-T cell populations
NK1.1/DX5Exclude or analyze NK cell populations
MHC I/IICharacterize APCs and context of antigen recognition
Isotype controlsControl for specificity in depletion or staining
Viability dyesExclude dead cells in cytometry panels

This structure reflects the typical experimental design in mouse immunology where clone 2.43 is used.

Clone 2.43 is a monoclonal antibody widely cited in scientific literature for its ability to deplete CD8+ T cells in vivo, mainly in mouse models. Key findings from studies using clone 2.43 include:

  • High specificity for CD8?: Clone 2.43 selectively targets the CD8? chain, resulting in specific depletion of CD8+ T cells without generally affecting other immune cell populations.
  • Established efficacy: Numerous studies across diverse research areas—such as immunology, aging, cancer, and infectious disease—have shown that 2.43 administration results in robust depletion of CD8+ T cells.
  • Versatility: The antibody is effective in multiple animal models, especially mice, and is frequently used to interrogate the role of CD8+ T cells in different biological contexts.

Representative applications and findings:

  • Aging and Immunology: Used in studies characterizing T cell memory populations and clonal expansion in aged versus young mice, demonstrating that clonally expanded CD8+ memory populations accumulate with age.
  • Infectious Disease: Clone 2.43-mediated CD8+ depletion allows researchers to test the contribution of CD8+ T cells in viral control; for example, CD8 depletion led to higher SARS-CoV-2 replication in mouse lungs, highlighting a key antiviral role for CD8+ T cells.
  • Cancer Models: Widely used to assess the role of CD8+ T cells in anti-tumor immunity, though specific cancer studies with clone 2.43 were not directly cited in the retrieved results.

Caveats and limitations:

  • Potential off-target or immunomodulatory effects: Although highly specific, repeated administration may induce immune responses against the antibody, compromising long-term experimental integrity or causing unintended immune effects.
  • Dosing variability: Effective depletion depends on dose, mouse strain, study design, and route of administration; common dosing ranges are 100–500 µg per mouse, with 250 µg as a typical standard dose.

In summary, clone 2.43 is a standard tool to dissect the function of CD8+ T cells in diverse in vivo studies, with findings relying on its specificity and effectiveness for targeted T cell depletion.

References & Citations

1.) Ardolino, M. et al. (2018) J Clin Invest. 128(10):4654-4668. PubMed
2.) Hawman DW, et al. (2021) Microorganisms 9(2):279 Journal Link
Depletion
FA
Flow Cytometry
ICC
IF Staining
IHC FFPE
in vivo Protocol
Immunoprecipitation Protocol

Certificate of Analysis

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